Fig. 5: I3P and I3A activate AHR signaling pathway to enhance TSG-6 production in MuSCs.

A The CYP1B1 expression in MuSCs stimulated with IFN-γ and TNF-α (10 ng/ml each) in the presence or absence of CH223191 (100 μM) for 24 h was measured by qRT-PCR. B The mRNA and protein levels of TSG-6 in MuSCs stimulated with IFN-γ and TNF-α (10 ng/ml each) in the presence or absence of CH223191 (100 μM) for 24 h were respectively measured by qRT-PCR and ELISA. C Immunofluorescence staining of AHR in MuSCs treated with IFN-γ (I, 10 ng/ml) and TNF-α (T, 10 ng/ml) in the presence or absence of I3P (50 μM) and I3A (100 μM) for 24 h. Scale bars, 40 μm. D MuSCs treated with IFN-γ (I, 10 ng/ml) and TNF-α (T, 10 ng/ml) in the presence or absence of I3P (50 μM), I3A (100 μM) and CH223191 (CH, 100 μM) were subjected to the cytoplasmic and nuclear extraction, and the distribution of AHR in cytoplasm and nucleus was analyzed by western blotting analysis. β-ACTIN and LAMINIB1 were served as loading controls for cytoplasmic and nuclear proteins, respectively. E The CYP1B1 expression in MuSCs stimulated with IFN-γ and TNF-α (10 ng/ml each) in the presence or absence of I3P (50 μM) and I3A (100 μM) for 24 h was measured by qRT-PCR. F The mRNA and protein levels of TSG-6 in MuSCs stimulated with IFN-γ and TNF-α (10 ng/ml each) in the presence or absence of I3P (50 μM) and CH223191 (CH, 100 μM) for 24 h were respectively measured by qRT-PCR and ELISA. G The mRNA and protein levels of TSG-6 in MuSCs stimulated with IFN-γ and TNF-α (10 ng/ml each) in the presence or absence of I3A (100 μM) and CH223191 (CH, 100 μM) for 24 h were respectively measured by qRT-PCR and ELISA. Data were shown as means ± SEM. Data were representative of three experiments with similar results. For two-group comparison, statistical analysis was performed by Student’s t test. For multiple group comparison, statistical analysis was performed by one-way ANOVA test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.