IL10 Antibody - #DF6894

FIGURE 4 Ox‐LDL/LPS activated ADAM17 to increase cleavage of mTIMD4 in macrophages. RAW264.7 cells treated with ox‐LDL (80 μg/mL) and TAPI‐1 (1 μM) for 24 h. (A) and (B) mTIMD4 expression was detected by immunofluorescent staining (n = 5). (C–F) The protein expression of mTIMD4, sTIMD4 in medium, TLR‐4, p‐NF‐κB, NF‐κB, IL‐6, caspase‐1, pro‐IL‐1β, TGF‐β1, and IL‐10 were detected by Western blotting (n = 3). (G–J) The mRNA expression mTIMD4, TLR‐4, IL‐6, and TGF‐β1 was determined by qRT‐PCR (n = 3–4). (K–N) RAW264.7 cells were treated with LPS (1 μg/mL) and TAPI‐1 (1 μM) for 24 h, and the protein expression of mTIMD4, sTIMD4 in medium, TLR‐4, p‐NF‐κB, NF‐κB, IL‐6, caspase‐1, pro‐IL‐1β, TGF‐β1, and IL‐10 were detected by Western blotting (n = 3). (O–P) RAW264.7 cells were transfected with 20 nM control si‐NC or si‐TIMD4 for 48 h and incubated with ox‐LDL (80 μg/mL) and TAPI‐1 (1 μM) for 24 h, followed by determination of mTIMD4, TLR‐4, p‐NF‐κB, NF‐κB, IL‐6, IL‐10, TGF‐β1, caspase‐1, pro‐IL‐1β protein expression by Western blotting (n = 3). HSP90 or GAPDH was used as a loading control. ns, not significant.

Fig. 3 IL-10 derived from hypoxic TAMs is required to maintain LVEM. a The different cytokines expression profiles among M0-N, M0-H, TAM-N and TAM-H were analysed by cytokine array (RayBio GSM-CAA-4000). b Screening and analysis of the differentially expressed cytokines. c The expressions of the five significant cytokines were analysed by qRT-PCR. d The secretions of the five significant cytokines were analysed by ELISA. e The migration effects of hypoxic TAMs-treated HDLECs on tumour cells (SiHa) and M2-polarized THP-1 macrophages were analysed by transwell assay in vitro. “Blank” represents the medium group. f Representative micrographs showing the tube formation in vitro (Scale bar, 50 μm). g Representative images showing the tube formation in vivo (Scale bar, 100 μm). h Statistical analysis showing the length of tube formation. Average length of tubes per field were calculated. i–l Popliteal lymphatic metastasis model was established in female C57BL/6 mice by inoculating the footpad with TC-1 cells (5 × 106). When footpad tumour size reached 50 mm3, IL-10 (50 ng/ml) or PBS was then injected into the centre of the tumours (n = 5/group, repeated twice) for 2 weeks daily. After 2 weeks of induction, primary tumours reached a comparable size of ~ 150 mm3, and then footpad tumours and popliteal LNs were collected for study. i Representative images of LYVE-1+ lymphatic vessel (red), CD206+ TAMs (green) and DAPI (blue) fluorescence staining in footpad tumour. j Metastasis-positive LNs were identified by IHC staining for epithelial marker CK7. k Statistical analysis showing the expression of peritumoural LV and LVEM in footpad tumour. l Statistical analysis showing the ratio of LNM. Error bars represent the mean ± SD of three independent experiments. **P < 0.01

Fig. 10: |Effects of XJEK on protein expression in MI mice with WB method(A-C) Representative figures of TNF-α, IL-1β and IL-10 protein. (D-F) Quantitative analyses of TNF-α, IL-1β and IL-10 protein. (mean ± S.D, n = 3).

Figure 7 MSC-ACE2 upregulates IL-10/STAT3/SOCS3 signaling pathway expression to suppress LPS-induced inflammation in EpH4-Ev cells. (A) Detection of relative protein expression levels of IL-10, phosphorylation levels of STAT3, STAT3 and SOCS3 by Western blot; (B–D), Statistics of the IL-10, phosphorylation levels of STAT3, STAT3 and SOCS3 Western blot results. Experiments were repeated three times and data were presented as the mean ± SEM (n = 4). * P < 0.05 vs. EpH4-Ev; # P < 0.05 vs. LPS; $ P < 0.05 vs. MSC; & P < 0.05 vs. MSC-GFP.

Figure S2. |Liver sections were IHC stained for TGF-β (a), IL-6 (b) and IL-10 (c). Three samples per group were examined, and representative images are shown (Scale bar represents 50 µm).