Lamin B1 Antibody - #AF5161

Fig. 7. NFATc3 promoted CPT2 transcription in gastrointestinal cancers. (A) The correlation between CPT2 and NFATc3 in GC and colon adenocarcinoma (COAD) from the GEPIA database. The r values and P values are from Pearson's correlation analysis. (B) The correlation between CPT2 and NFATc3 in GC (n = 36) and CRC (n = 55) samples. The r values and P values are from Pearson's correlation analysis. (C) CPT2 levels were downregulated when NFATc3 was knocked down in the indicated cells. Error bars, SD of three independent experiments. *P < 0.05 or **P < 0.01 versus the control. (D) Western blot analysis shows the NFATc3 content in the cytoplasm and nucleus of HGC27 cells treated with or without oxaliplatin (Oxa, 40 μM) for 24 h. Lamin B was used as the nuclear loading control with α- Tubulin as the cytosolic loading control. (E) NFATc3 DNA-binding sites are present in the human CPT2 promoter region. (F) ChIP-PCR in HGC27 and HCT116 cells. The data are representative of two independent experiments. *P < 0.05 or **P < 0.01. (G) Relative CPT2 luciferase promoter activity in the indicated cells with NFATc3 depletion or overexpression. Error bars, SD of three independent experiments. *P < 0.05 or **P < 0.01 versus the control. (H) Proposed working model from this study. Oxaliplatin-induced ROS generation causes the accumulation of NFATc3 in the nucleus, promoting the CPT2 transcription. Perhexiline, as a CPT inhibitor, blocks FA catabolism in mitochondria, resulting in reduced NADPH levels. Therefore, increased ROS caused the apoptosis in gastrointestinal cancers.

Fig. 3. PHI treatment reduced M1 macrophage-induced HSC activation. (a) mHSCs were co-cultured with the CMs from RAW264.7 cells with LPS/IFNγ and PHI treatment for 12 h. (b-g) The expression of MMP2, TIMP1, TGF-β, α-SMA, COL1 and NF-κB mRNA in mHSCs after co-culture with macrophage-derived CMs for 24 h was detected by RT-qPCR (n = 3). (h) The expression of TGF-β, α-SMA, COL1, P65 and P-P65 proteins in mHSCs after co-culture with macrophage-derived CMs for 24 h was detected by western blotting. (i) The relative quantification of TGF-β, α-SMA, COL1 and P-P65/P65 protein expression in western blotting results was analyzed by ImageJ software (n = 3). (j) The expression of cytoplasm and nucleus NF-κB P65 proteins in mHSCs after co-culture with macrophage-derived CMs for 24 h was detected by western blotting. (k, l) The relative quantification of cytoplasm and nucleus NF-κB P65 protein expression in western blotting results was analyzed by ImageJ software (n = 3). (m) Immunofluorescence staining of α-SMA in mHSCs after co-culture with CMs from RAW264.7 cells with different treatments for 24 h (Scale bar=100 µm). Results are presented as mean ± SD. ###P 

Fig. 6. The impact of ellagic acid treatment on arsenic-induced Nrf2/Keap1 signaling pathway in the hippocampus and HT22 cells. (A) Western blot for p-Nrf2, Nrf2 and Keap1 in the hippocampus. (B-D) Relative density analysis of the p-Nrf2, Nrf2 and Keap1 protein bands in the hippocampus. (E) Quantification of Keap1 mRNA levels in the hippocampus. (F) Western blot for Nucleus Nrf2, Nrf2 and Keap1 in HT22 cells. (G-I) Relative density analysis of the Nucleus Nrf2, Nrf2 and Keap1 protein bands in HT22 cells. (J) Quantification of Keap1 mRNA levels in HT22 cells. Data are presented as mean ± SD (n=3). * significant difference compared to the control group (*P

FIGURE 6 Downregulation of microRNA‐150‐5p reduced NF‐κB mediated cardiac inflammation. (a) CCK‐8 assay indicated that AMO‐150‐5p transfection decreased CFs viability compared with HG treatment. *p < .05 versus control. # p < .05 versus HG. (b,c) Western blot results showed that AMO‐150‐5p reduced the nuclear phosphorylated NF‐κB p65 subunit (Nu‐p65) level but increased the cytoplasmic NF‐κB p65 subunit (Cyto‐p65) level in HG treated CFs. *p < .05 versus control. # p < .05 versus HG. Three times independent in each group. (d) CFs were transfected with pNF‐κB‐Luc reporter plasmid and AMO‐150‐5p under HG treatment. The dual‐luciferase reporter assay was performed to compare NF‐κB activity. n = 3, *p < .05 versus control. #p < .05 versus HG. (E) AMO‐150‐5p suppressed IL‐1β production induced by HG treatment by ELISA detection. n = 3, *p < 0.05 versus control. #p < .05 versus HG. (f) ELISA results showed that NF‐κB agonist (LPS) reversed the protective role of AMO‐150‐5p by an increase in IL‐1β. n = 3, *p < .05 versus control. #p < .05 versus HG. $p < .05 versus AMO‐150‐5p. AMO‐150‐5p: miR‐150‐5p inhibitor. CCK‐8, cell counting kit‐8; CFs, cardiac fibroblasts; ELISA, enzyme linked immunosorbent assay; HG, high glucose