Fig. 6.

Supernatant containing Dectin-1/Syk inhibited macrophages alleviated renal fibrosis in vitro. SV40 cells were incubated with the supernatants from the RAW cells fellow the approach described in Figs. 4, 5. A SV40 cells were treated with the supernatants from the RAW cells fellow the approach described in Fig. 5A for 6 h. Total proteins were extracted and subjected to analysis of p-Smad3 and Smad3 protein levels. GAPDH was used as loading control; B Western blot detected Smad3, Lamin B and GAPDH protein levels in the nuclear extractions and cytoplasm. C Representative images of Smad3 nuclear translocation detected by immune fluorescence microscopy; D Representative western blot analysis of COL1α1, COL4α2 and α-SMA of each group in SV40 cells. GAPDH was used as loading control; E Real-time qPCR showing mRNA levels of Col4a2, Col1a1 and Acta2 of each group in SV40 cells.; F SV40 cells were treated with the supernatants from the RAW cells fellow the approach described in Fig. 5E for 6 h. Total proteins were extracted and subjected to analysis of p-Smad3 and Smad3 protein levels. GAPDH was used as loading control; G Western blot detected Smad3, Lamin B and GAPDH protein levels in the nuclear extractions and cytoplasm. H Representative images of Smad3 nuclear translocation detected by immune fluorescence microscopy; I Representative western blot analysis of COL1α1, COL4α2 and α-SMA of each group in SV40 cells. GAPDH was used as loading control; [A-I, n = 3; 1-way ANOVA followed by Tukey post-hoc tests (A-J: number of comparisons = 6; *P < 0.05, **P < 0.01, and ***P < 0.001]