IL10 Antibody - #DF6894

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).

Figure 5. ARR downregulates the protein expression of TRAF6 and NF-κB p65 by increasing the content of NLRC3 protein molecules. (A) Expression levels of NLRC3, TRAF6 and NF-κB p65 in LPS-stimulated RAW264.7 cells were detected using immunohistochemistry staining (magnification, ×400; scale bar, 200-µm). The mean optical density values of (B) NLRC3, (C) TRAF6 and (D) NF-κB p65 were quantified using ImageJ software. The protein expression levels of (E) NLRC3 and (F) TRAF6 were detected using western blotting and semi-quantified using Image Lab software. LPS represents protein from the 100 ng/ml LPS-treated group; Indo represents protein from the 8 µg/ml Indo and 100 ng/ml LPS-treated group; ARR represents protein from the 100 µg/ml ARR and 100 ng/ml LPS-treated group. Data are presented as the mean ± SD (n=3). ##P<0.01 vs. control group; *P<0.05, **P<0.01 vs. LPS group. ARR, α-rhamnrtin-3-α-rhamnoside; TRAF6, tumor necrosis factor-associated factor 6; NLRC3, NOD-like receptor family CARD domain containing 3; LPS, lipopolysaccharide; Indo, indomethacin.

FIGURE 2 Phenotypic characteristics of macrophages in EtOH-fed mice liver tissue and ethanol-stimulated RAW264.7 cells. (A) Effects of ethanol on protein and mRNA levels of M1 macrophage markers (TNF-α, IL-1β) in liver tissue. (B) The protein and mRNA levels of M2 macrophage markers (IL-10, ARG-1) in liver tissue. (C) Immunofluorescence staining results of M1, M2 macrophage markers (CD86, CD206) in liver tissue of EtOH-fed mice. (D) The protein and mRNA levels of M1 macrophage (TNF-α, IL-1β) in ethanol-induced RAW264.7 cells. (E) The protein and mRNA levels of M2 macrophage markers (IL-10, ARG-1) in ethanol-induced RAW264.7 cells. The results are shown as relative expression against control expression without treatment. Data shown are the mean ± SD from three independent experiments. *p < 0.05, **p < 0.01 vs. CD-fed group or control group.

Fig. 4. |TUDCA treatment suppressed LPS-activated NF-κB signaling and the production of proinflammatory cytokines. (A) Representative immunoblots of the nuclear NF-κB p65 in the hippocampus was shown, and Histone H3 was used as a loading control. (B) Representative immunoblots of TNF-α, IL-1β, IL-6 and IL-10 in the hippocampus, and β-actin protein was used here as an internal control.

FIGURE 5 | WMW reduces colonic inflammatory responses in DSS-induced colitis. (B) Relative mRNA level of IL10, Arg-1, and TGF-β1in the colon was measured through qPCR. Expression of IL-10, Arg-1, and TGF-β1 in colon tissue lysates were determined by Western blot from DSS-challenged mice. n = 3,.means ± SEM.*p < 0.05, **p < 0.01, ***p < 0.001.

FIGURE 6 | The proportion of M1/M2 macrophages changes in colon sections obtained from mice with ulcerative colitis induced by DSS (A) Double immunofluorescent signals from CD80 and interleukin-1 in DSS-induced colitis tissue at different time points (original magnification, 200 ×). (B) Double immunofluorescent signals from CD206 and interleukin-10 in DSS-induced colitis tissue at different time points (original magnification, 200 ×).