Phospho-STAT3 (Tyr705) Antibody | Affinity Biosciences LTD|亲科生物官网

产品:	磷酸化 STAT3 (Tyr705) 抗体
货号:	AF3293
描述:	Rabbit polyclonal antibody to Phospho-STAT3 (Tyr705)
应用:	WB IHC IF/ICC IP
反应:	Human, Mouse, Rat
预测:	Pig, Zebrafish, Bovine, Horse, Sheep, Rabbit, Chicken
分子量:	86kDa; 88kD(Calculated).
蛋白号:	P40763
RRID:	AB_2810278

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阻断肽(封闭肽)是特异性结合目标抗体和阻断抗体结合的多肽。这些多肽包含抗体识别的抗原表位。与阻断肽结合的抗体不再与靶蛋白上的表位结合。例如，在免疫印迹(WB)和免疫组化(IHC)中，通过比较被阻断抗体和未阻断抗体的染色，可以看到哪种染色是特异性的。

规格	价格	库存
50ul	RMB¥ 1300	现货
100ul	RMB¥ 2400	现货
200ul	RMB¥ 3200	现货

货期: 当天发货

联系销售

实验方案

产品描述

来源:

Rabbit

应用:

WB 1:500-1:2000, IHC 1:50-1:200, IP, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: 适用于变性蛋白样本的免疫印迹检测. IHC: 适用于组织样本的石蜡(IHC-p)或冰冻(IHC-f)切片样本的免疫组化/荧光检测. IF/ICC: 适用于细胞样本的荧光检测. ELISA(peptide): 适用于抗原肽的ELISA检测.

反应:

Human,Mouse,Rat

预测:

Pig(100%), Zebrafish(100%), Bovine(100%), Horse(100%), Sheep(91%), Rabbit(100%), Chicken(100%)

克隆:

Polyclonal

特异性:

Phospho-STAT3 (Tyr705) Antibody detects endogenous levels of STAT3 only when phosphorylated at Tyrosine 705.

RRID:

AB_2810278
引用格式: Affinity Biosciences Cat# AF3293, RRID:AB_2810278.

偶联:

Unconjugated.

纯化:

The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.

保存:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

别名:

展开/折叠

1110034C02Rik; Acute Phase Response Factor; Acute-phase response factor; ADMIO; APRF; AW109958; DNA binding protein APRF; FLJ20882; HIES; MGC16063; Signal transducer and activator of transcription 3 (acute phase response factor); Signal transducer and activator of transcription 3; STAT 3; Stat3; STAT3_HUMAN;

抗原和靶标

免疫原:

Uniprot:

P40763(STAT3_HUMAN) >>Visit HPA database.

基因/基因ID:

STAT3(6774)

表达:

P40763 STAT3_HUMAN:

Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.

描述:

The protein encoded by this gene is a member of the STAT protein family. In response to cytokines and growth factors, STAT family members are phosphorylated by the receptor associated kinases, and then form homo- or heterodimers that translocate to the cell nucleus where they act as transcription activators.

序列:

P40763 STAT3_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MAQWNQLQQLDTRYLEQLHQLYSDSFPMELRQFLAPWIESQDWAYAASKESHATLVFHNLLGEIDQQYSRFLQESNVLYQHNLRRIKQFLQSRYLEKPMEIARIVARCLWEESRLLQTAATAAQQGGQANHPTAAVVTEKQQMLEQHLQDVRKRVQDLEQKMKVVENLQDDFDFNYKTLKSQGDMQDLNGNNQSVTRQKMQQLEQMLTALDQMRRSIVSELAGLLSAMEYVQKTLTDEELADWKRRQQIACIGGPPNICLDRLENWITSLAESQLQTRQQIKKLEELQQKVSYKGDPIVQHRPMLEERIVELFRNLMKSAFVVERQPCMPMHPDRPLVIKTGVQFTTKVRLLVKFPELNYQLKIKVCIDKDSGDVAALRGSRKFNILGTNTKVMNMEESNNGSLSAEFKHLTLREQRCGNGGRANCDASLIVTEELHLITFETEVYHQGLKIDLETHSLPVVVISNICQMPNAWASILWYNMLTNNPKNVNFFTKPPIGTWDQVAEVLSWQFSSTTKRGLSIEQLTTLAEKLLGPGVNYSGCQITWAKFCKENMAGKGFSFWVWLDNIIDLVKKYILALWNEGYIMGFISKERERAILSTKPPGTFLLRFSESSKEGGVTFTWVEKDISGKTQIQSVEPYTKQQLNNMSFAEIIMGYKIMDATNILVSPLVYLYPDIPKEEAFGKYCRPESQEHPEADPGSAAPYLKTKFICVTPTTCSNTIDLPMSPRTLDSLMQFGNNGEGAEPSAGGQFESLTFDMELTSECATSPM

种属预测

种属预测:

score>80的预测可信度较高，可尝试用于WB检测。*预测模型主要基于免疫原序列比对，结果仅作参考，不作为质保凭据。

Species

Results

Score

Pig

100

Horse

100

Bovine

100

Zebrafish

100

Chicken

100

Rabbit

100

Sheep

Dog

Xenopus

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

翻译修饰 - P40763 作为底物

P40763

Site	PTM Type	Enzyme	Source
A2	Acetylation		Uniprot
S25	Phosphorylation		Uniprot
Y45	Phosphorylation		Uniprot
K49	Acetylation		Uniprot
K49	Methylation		Uniprot
Y68	Phosphorylation		Uniprot
Y79	Phosphorylation		Uniprot
K87	Acetylation		Uniprot
K87	Ubiquitination		Uniprot
K97	Ubiquitination		Uniprot
K140	Ubiquitination		Uniprot
K153	Ubiquitination		Uniprot
K161	Ubiquitination		Uniprot
K163	Ubiquitination		Uniprot
Y176	Phosphorylation		Uniprot
K177	Sumoylation		Uniprot
K177	Ubiquitination		Uniprot
K180	Ubiquitination		Uniprot
S181	Phosphorylation		Uniprot
S194	Phosphorylation		Uniprot
T196	Phosphorylation		Uniprot
K199	Ubiquitination		Uniprot
T236	Phosphorylation		Uniprot
K244	Ubiquitination		Uniprot
K283	Ubiquitination		Uniprot
K290	Ubiquitination		Uniprot
K294	Ubiquitination		Uniprot
K318	Ubiquitination		Uniprot
K348	Ubiquitination		Uniprot
K363	Ubiquitination		Uniprot
K365	Ubiquitination		Uniprot
K370	Acetylation		Uniprot
K370	Ubiquitination		Uniprot
K383	Ubiquitination		Uniprot
K409	Ubiquitination		Uniprot
S429	Phosphorylation		Uniprot
T440	Phosphorylation		Uniprot
Y446	Phosphorylation		Uniprot
K451	Sumoylation		Uniprot
K495	Sumoylation		Uniprot
Y539	Phosphorylation		Uniprot
S540	Phosphorylation		Uniprot
K551	Ubiquitination		Uniprot
K574	Ubiquitination		Uniprot
K601	Acetylation		Uniprot
K601	Ubiquitination		Uniprot
T605	Phosphorylation		Uniprot
K615	Acetylation		Uniprot
K615	Ubiquitination		Uniprot
K626	Ubiquitination		Uniprot
K631	Acetylation		Uniprot
K631	Ubiquitination		Uniprot
T632	Phosphorylation		Uniprot
S636	Phosphorylation		Uniprot
Y640	Phosphorylation		Uniprot
T641	Phosphorylation		Uniprot
K642	Ubiquitination		Uniprot
Y674	Phosphorylation		Uniprot
K679	Ubiquitination		Uniprot
K685	Acetylation		Uniprot
K685	Ubiquitination		Uniprot
Y686	Phosphorylation		Uniprot
S691	Phosphorylation		Uniprot
S701	Phosphorylation		Uniprot
Y705	Phosphorylation	O60674 (JAK2) , Q15300 (RET/PTC2) , P23458 (JAK1) , P22455 (FGFR4) , P27361 (MAPK3) , Q13882 (PTK6) , P22607 (FGFR3) , Q9UM73 (ALK) , P14618 (PKM) , P16591 (FER) , P12931 (SRC)	Uniprot
K707	Acetylation		Uniprot
K707	Ubiquitination		Uniprot
T708	Phosphorylation		Uniprot
T714	Phosphorylation		Uniprot
T717	Phosphorylation		Uniprot
S719	Phosphorylation		Uniprot
T721	Phosphorylation		Uniprot
S727	Phosphorylation	P51812 (RPS6KA3) , P45984-2 (MAPK9) , Q13233 (MAP3K1) , O75582 (RPS6KA5) , P28482 (MAPK1) , Q13555 (CAMK2G) , O43293 (DAPK3) , P42345 (MTOR) , P27361 (MAPK3) , Q02156 (PRKCE) , P45983 (MAPK8) , P51617 (IRAK1) , Q16539 (MAPK14) , Q05655 (PRKCD) , Q9UBE8 (NLK) , P06493 (CDK1) , Q00535 (CDK5) , Q9HC98 (NEK6)	Uniprot
S754	Phosphorylation		Uniprot

研究背景

P40763_STAT3_HUMAN

功能:

Signal transducer and transcription activator that mediates cellular responses to interleukins, KITLG/SCF, LEP and other growth factors. Once activated, recruits coactivators, such as NCOA1 or MED1, to the promoter region of the target gene. May mediate cellular responses to activated FGFR1, FGFR2, FGFR3 and FGFR4. Binds to the interleukin-6 (IL-6)-responsive elements identified in the promoters of various acute-phase protein genes. Activated by IL31 through IL31RA. Acts as a regulator of inflammatory response by regulating differentiation of naive CD4(+) T-cells into T-helper Th17 or regulatory T-cells (Treg): deacetylation and oxidation of lysine residues by LOXL3, leads to disrupt STAT3 dimerization and inhibit its transcription activity. Involved in cell cycle regulation by inducing the expression of key genes for the progression from G1 to S phase, such as CCND1. Mediates the effects of LEP on melanocortin production, body energy homeostasis and lactation (By similarity). May play an apoptotic role by transctivating BIRC5 expression under LEP activation. Cytoplasmic STAT3 represses macroautophagy by inhibiting EIF2AK2/PKR activity. Plays a crucial role in basal beta cell functions, such as regulation of insulin secretion (By similarity).

翻译修饰:

Tyrosine phosphorylated upon stimulation with EGF. Tyrosine phosphorylated in response to constitutively activated FGFR1, FGFR2, FGFR3 and FGFR4 (By similarity). Activated through tyrosine phosphorylation by BMX. Tyrosine phosphorylated in response to IL6, IL11, LIF, CNTF, KITLG/SCF, CSF1, EGF, PDGF, IFN-alpha, LEP and OSM. Activated KIT promotes phosphorylation on tyrosine residues and subsequent translocation to the nucleus. Phosphorylated on serine upon DNA damage, probably by ATM or ATR. Serine phosphorylation is important for the formation of stable DNA-binding STAT3 homodimers and maximal transcriptional activity. ARL2BP may participate in keeping the phosphorylated state of STAT3 within the nucleus. Upon LPS challenge, phosphorylated within the nucleus by IRAK1. Upon erythropoietin treatment, phosphorylated on Ser-727 by RPS6KA5. Phosphorylation at Tyr-705 by PTK6 or FER leads to an increase of its transcriptional activity. Dephosphorylation on tyrosine residues by PTPN2 negatively regulates IL6/interleukin-6 signaling.

Acetylated on lysine residues by CREBBP. Deacetylation by LOXL3 leads to disrupt STAT3 dimerization and inhibit STAT3 transcription activity. Oxidation of lysine residues to allysine on STAT3 preferentially takes place on lysine residues that are acetylated.

Some lysine residues are oxidized to allysine by LOXL3, leading to disrupt STAT3 dimerization and inhibit STAT3 transcription activity. Oxidation of lysine residues to allysine on STAT3 preferentially takes place on lysine residues that are acetylated.

(Microbial infection) Phosphorylated on Tyr-705 in the presence of S.typhimurium SarA.

S-palmitoylated by ZDHHC19 in SH2 putative lipid-binding pockets, leading to homodimerization. Nuclear STAT3 is highly palmitoylated (about 75%) compared with cytoplasmic STAT3 (about 20%).

S-stearoylated, probably by ZDHHC19.

细胞定位:

Cytoplasm. Nucleus.
Note: Shuttles between the nucleus and the cytoplasm. Translocated into the nucleus upon tyrosine phosphorylation and dimerization, in response to signaling by activated FGFR1, FGFR2, FGFR3 or FGFR4. Constitutive nuclear presence is independent of tyrosine phosphorylation. Predominantly present in the cytoplasm without stimuli. Upon leukemia inhibitory factor (LIF) stimulation, accumulates in the nucleus. The complex composed of BART and ARL2 plays an important role in the nuclear translocation and retention of STAT3. Identified in a complex with LYN and PAG1.

组织特异性:

Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.

亚基结构:

Forms a homodimer or a heterodimer with a related family member (at least STAT1). Interacts with IL31RA, NCOA1, PELP1, SIPAR, SOCS7, STATIP1 and TMF1 (By similarity). Interacts with IL23R in presence of IL23. Interacts (via SH2 domain) with NLK. Interacts with ARL2BP; the interaction is enhanced by LIF and JAK1 expression (By similarity). Interacts with KPNA4 and KPNA5; KPNA4 may be the primary mediator of nuclear import (By similarity). Interacts with CAV2; the interaction is increased on insulin-induced tyrosine phosphorylation of CAV2 and leads to STAT3 activation (By similarity). Interacts with ARL2BP; interaction is enhanced with ARL2. Interacts with NEK6 (By similarity). Binds to CDK9 when activated and nuclear. Interacts with BMX. Interacts with ZIPK/DAPK3. Interacts with PIAS3; the interaction occurs on stimulation by IL6, CNTF or OSM and inhibits the DNA binding activity of STAT3. In prostate cancer cells, interacts with STAT3 and promotes DNA binding activity of STAT3. Interacts with STMN3, antagonizing its microtubule-destabilizing activity. Interacts with the 'Lys-129' acetylated form of BIRC5/survivin. Interacts with FER. Interacts (via SH2 domain) with EIF2AK2/PKR (via the kinase catalytic domain). Interacts with STAT3; the interaction is independent of STAT3 Tyr-705 phosphorylation status. Interacts with FGFR4. Interacts with OCAD1 (By similarity). Interacts with ZDHHC19, leading to palmitoylation which promotes homodimerization and activation.

(Microbial infection) Interacts with HCV core protein.

(Microbial infection) Interacts with S.typhimurium SarA.

蛋白家族:

Belongs to the transcription factor STAT family.

研究领域

· Cellular Processes > Cell growth and death > Necroptosis. (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Signaling pathways regulating pluripotency of stem cells. (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > FoxO signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > Jak-STAT signaling pathway. (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.

· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Measles.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer. (View pathway)

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Pancreatic cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Acute myeloid leukemia. (View pathway)

· Human Diseases > Cancers: Specific types > Non-small cell lung cancer. (View pathway)

· Human Diseases > Immune diseases > Inflammatory bowel disease (IBD).

· Organismal Systems > Immune system > Chemokine signaling pathway. (View pathway)

· Organismal Systems > Immune system > Th17 cell differentiation. (View pathway)

· Organismal Systems > Endocrine system > Prolactin signaling pathway. (View pathway)

· Organismal Systems > Endocrine system > Adipocytokine signaling pathway.

文献引用

1). Gut dysbiosis promotes prostate cancer progression and docetaxel resistance via activating NF-κB-IL6-STAT3 axis. Microbiome, 2022 (PubMed: 35710492) [IF=15.5]

Application: IF/ICC Species: Mouse Sample: tumor tissue

Fig. 3 Intratumoral LPS activated NF-κB-IL6-STAT3 axis. A LPS levels in mouse feces and serum by ELISA; HE staining (scale bar, 50 μm) and histology score for colon tissue in the Abx and NC group. B Immunohistochemistry (scale bar, 50 μm) for LPS in subcutaneous and orthotopic tumor tissues and western blot of intratumoral LPS levels from three biological duplications for the Abx and NC group. C Transcription levels of cytokines by RT-qPCR and protein levels of IL6 in cell supernatant by ELISA in RM-1 cultured with LPS (100 μg/ml) for 24 h. D, E Immunofluorescence (scale bar, 100 μm) for p-p65 and p-STAT3 in RM-1 cultured with or without LPS for 24 h; western blot of relative proteins for RM-1 cultured with LPS at different concentrations for 24 h; transcription levels of IL6 by RT-qPCR and western blot of relative proteins in RM-1 cultured with LPS or LPS with BAY-11-7082 for 24 h; protein levels of p-STAT3 and STAT3 in RM-1 cultured with CM or CM with antibody-IL6 for 24 h. F, G IL6 levels in tumor tissue lysate and serum by ELISA. Western blot of relative proteins in tumor from three biological duplications. Immunohistochemistry of tumor tissues for p-p65- and p-STAT3-positive cell (scale bar, 50 μm). Statistical significance was assessed by unpaired Student’s T-test or LSD in one-way ANOVA. *p < 0.05, **p < 0.01, and ***p < 0.001: compared to the NC group; #p < 0.05, ##p < 0.01, and ###p < 0.001: compared to the LPS or CM group

Application: WB Species: Mouse Sample: tumor tissue

Application: IHC Species: Mouse Sample: tumor tissue

2). A Bioinspired Manganese-Organic Framework Ameliorates Ischemic Stroke through its Intrinsic Nanozyme Activity and Upregulating Endogenous Antioxidant Enzymes. Advanced Science, 2023 (PubMed: 37129343) [IF=15.1]

Application: WB Species: Mouse Sample: N2a cells

Figure 3 The effect of pDA-MNOF on upregulating HO1 and SOD2 via STAT3 signaling. a) Western blot analysis of p-STAT3, STAT3, HO1, and SOD2 expression in N2a cells treated with 12.5 µg mL−1 pDA-MNOF for various time durations. b) Quantification of the band intensity ratios of p-STAT3/STAT3, HO1/β-actin, and SOD2/β-actin in (a). t-STAT3 indicated total proteins of p-STAT3 and STAT3. c) Quantification of fluorescent intensity of p-STAT3, HO1, and SOD2 in PBS or 12.5 µg mL−1 pDA-MNOF-treated N2a cells (n = 4). Data were presented with mean ± s.d.; *, p < 0.05; **, p < 0.01; T-test. d) Representative images of immunofluorescent staining for p-STAT3, HO-1, and SOD-2 (red) in indicated groups. Nuclei were stained in blue, and cell body was outlined with white dashed lines. Scale bar, 20 µm. e) Western blot analysis of p-STAT3, STAT3, HO1, and SOD2 expression in N2a cells treated with different concentrations of pDA-MNOF for 12 h. f) Quantification of the band intensity ratios of p-STAT3/STAT3, HO1/β-actin, and SOD2/β-actin in (e). g) Experimental scheme for the RNA interference and pathway inhibition assays. h) Western blot analysis of p-STAT3 and STAT3 in N2a cells treated with PBS, siRNA Control (siRNA Ctrl), siRNA 1#, siRNA 2#, and siRNA 3#. i) Quantification of the band intensity ratios of p-STAT3/β-actin and STAT3/β-actin in (h). j) N2a cells were transfected with siRNA Ctrl, siRNA 2#, and siRNA 3#, followed by treatment with pDA-MNOF; the lysates were analyzed with indicated antibodies. k) Quantification of the band intensity ratios of p-STAT3/β-actin, HO1/β-actin, and SOD2/β-actin in (j). Data were presented with mean ± s.d.; *, p < 0.05; ANOVA. l) N2a cells were treated with PBS or pDA-MNOF, followed by treatment with AG490; the lysates were analyzed with indicated antibodies. m) Quantification of the band intensity ratios of p-STAT3/STAT3, HO1/β-actin, and SOD2/β-actin in (l) (n = 3).

3). Exosomal circPOLQ promotes macrophage M2 polarization via activating IL-10/STAT3 axis in a colorectal cancer model. Journal for immunotherapy of cancer, 2024 (PubMed: 38782541) [IF=10.9]

Application: WB Species: Human Sample:

Figure 7 Exosomal circPOLQ induces macrophage M2 polarization through the mir-379–3 p/IL-10/STAT3 axis. (A) TargetScan Human V.7.2 database predicts that miR-379–3 p may target the binding site of IL-10 and the mutation site of IL-10 MT. (B) A luciferase reporter gene activity assay was performed to determine the effect of the miR-379–3 p mimic on the 3’-UTR luciferase activity of WT/MT IL-10. (C) RNA pull-down was performed in 293 T cells, followed by qPCR and nucleic acid electrophoresis to detect the enrichment of IL-10. (D) Macrophages treated with HCT116 or SW620 exosomes were co-transfected with miR-379–3 p mimics and IL-10-overexpressing plasmids, and the expression of macrophage M2 polarization markers was detected by qPCR. (E) Macrophages treated with HCT116 or SW620 exosomes were co-transfected with miR-379–3 p inhibitor and IL-10 siRNA, and the expression of macrophage M2 polarization markers was detected by qPCR. (F) Macrophages treated with HCT116 or SW620 exosomes were transfected with circPOLQ siRNA, and the expressions of p-STAT3 and STAT3, key molecules of STAT3 signaling pathway, were detected by western blot. (G) Macrophages treated with HCT116 or SW620 exosomes were co-transfected with circPOLQ siRNA and miR-379–3 p inhibitor, and the expressions of p-STAT3 and STAT3, key molecules of STAT3 signaling pathway, were detected by western blot. (H) Macrophages treated with HCT116 exosomes were co-transfected with miR-379–3 p mimics and overexpressed IL-10 plasmids, and the key molecules of STAT3 signaling pathway p-STAT3 and macrophage M2 polarization markers were detected by western blot. (I) Macrophages treated with HCT116 exosomes were co-transfected with miR-379–3 p inhibitor and IL-10 siRNA, and the key molecules of STAT3 signaling pathway p-STAT3 and macrophage M2 polarization markers were detected by western blot. (J, K) Macrophages treated with HCT116 exosomes were co-transfected with circPOLQ siRNA and overexpressed IL-10 plasmids, and the expression of macrophage M2 polarization markers was detected by qPCR. (L, M) Macrophages treated with HCT116 exosomes were co-transfected with circPOLQ siRNA and overexpressed IL-10 plasmids, and the expression of CD206 was detected by FACS and the positive rate of CD206 was measured by FlowJo. Statistical significance was calculated by Student’s t-test. Data in the text were expressed as mean±SD, *p

4). Aligned electrospun poly(l-lactide) nanofibers facilitate wound healing by inhibiting macrophage M1 polarization via the JAK-STAT and NF-κB pathways. JOURNAL OF NANOBIOTECHNOLOGY, 2022 (PubMed: 35883095) [IF=10.2]

Application: WB Species: Mice Sample:

Fig. 3 The underlying mechanism by which aligned fibers affected macrophage polarization. A Venn diagram showing differentially expressed genes. B KEGG pathway analysis between the A20 and R20 groups. C Heatmap of differentially expressed genes among the three groups. D Heatmap of macrophage polarization-related genes between the A20 and R20 groups. E Volcano diagram of differentially expressed genes. F Western blot analysis of the NF-κB signaling pathway. G Immunofluorescence staining showing the nuclear translocation of NF-κB p65. The nucleus is stained blue, and NF-κB p65 protein is stained red. H Western blot images and semiquantitative analysis of the JAK-STAT signaling pathway (*p < 0.05, **p < 0.01, n = 3)

5). Tofacitinib Promotes Functional Recovery after Spinal Cord Injury by Regulating Microglial Polarization via JAK/STAT Signaling Pathway. International Journal of Biological Sciences, 2023 (PubMed: 37781508) [IF=9.2]

6). P21 and P27 promote tumorigenesis and progression via cell cycle acceleration in seminal vesicles of TRAMP mice. International Journal of Biological Sciences, 2019 (PubMed: 31592235) [IF=9.2]

7). Protopanaxadiol inhibits epithelial-mesenchymal transition of hepatocellular carcinoma by targeting STAT3 pathway. Cell Death & Disease, 2019 (PubMed: 31431619) [IF=9.0]

Application: WB Species: human Sample: PLC/PRF/5 cells

Fig. 4| PPD inhibited EMT of HCC cells by targeting STAT3. a Molecular docking scores for PPD and six potential targets. b Predicted model of PPD binding to the SH2 domain of STAT3 as shown by computational docking. The protein backbone is portrayed as a transparent green ribbon, and PPD is depicted as atom-red sticks-and-balls. c Biacore analysis results of PPD and STAT3. d, e PPD inhibited the translocation of STAT3 to the nucleus in PLC/PRF/5 cells detected by immunofluorescence and Western blot

8). DINP aggravates autoimmune thyroid disease through activation of the Akt/mTOR pathway and suppression of autophagy in Wistar rats. ENVIRONMENTAL POLLUTION, 2019 (PubMed: 30447474) [IF=8.9]

9). Acid sphingomyelinase downregulation alleviates vascular endothelial leptin resistance in rats. ACTA PHARMACOLOGICA SINICA, 2020 (PubMed: 31848475) [IF=8.2]

Application: WB Species: Rat Sample: rat aortic endothelial cells (RAECs)

Fig. 1 ASM downregulation improved PA-induced leptin resistance in RAECs. Western blot gels and summarized data showing the protein expression of Ob-Rb (a) and SOCS3 (b) in RAECs treated with PA (0.1, 0.2, and 0.3 mM) with or without 100 nM leptin. RAECs were incubated with 0.3 mM PA for 24 h and were then transfected with ASM siRNA for 24 h. Then, 100 nM leptin was added for 15 min prior to the collection of the cells. Then, Western blotting and an AmpliteTM Fluorimetric Kit were used to test the protein expression of ASM (c) and ASM activity (d), respectively, in RAECs. Representative fluorescence images (e, scale bar = 20 μm) and summarized data showing the mean fluorescent intensity (MFI) of a FITC-labeled anti-ceramide antibody (f). The photographs were taken at ×400 magnification. The protein expression of ObRb (g) and SOCS3 (h) and the ratio of p-STAT3/STAT3 (i) was determined using Western blotting. The data are the means ± SEMs from three experiments. *P < 0.05 vs. scramble control (Scr Ctrl); #P < 0.05 vs. the PA alone-treated group

10). Formononetin protects against inflammation associated with cerebral ischemia-reperfusion injury in rats by targeting the JAK2/STAT3 signaling pathway. BIOMEDICINE & PHARMACOTHERAPY, 2022 (PubMed: 35339827) [IF=7.5]

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