Fig. 5: Prom1 deficiency enhanced TGFβ signaling in hepatocytes.
From: Hepatocyte-specific Prominin-1 protects against liver injury-induced fibrosis by stabilizing SMAD7
A–E Primary hepatocytes (PHs) were isolated from 8-week-old male Prom1+/+ (WT) and Prom1−/− (KO) mice, grown for 4 h, serum-starved for 16 h, and treated with 5 ng/ml TGFβ for the indicated times. The cells were analyzed by immunofluorescence analysis of p-SMAD2/3 and SMAD7 (A); immunoblot analysis of PROM1, p-SMAD2/3, SMAD2/3, SMAD7, cleaved Caspase-3 and GAPDH (C); and RT–qPCR analysis of the Smad7 and Prom1 mRNA levels. Each mRNA level was normalized to that of 18 S rRNA (E). B The percentage of cells with nuclear p-SMAD2/3 was statistically determined (n = 3 for each group). D The band intensities of p-SMAD2/3, SMAD7, and cleaved Caspase-3 in (C) were statistically analyzed after normalization to the intensity of GAPDH (n = 3 for each group). F, G Primary KO hepatocytes were infected by adenovirus expressing LacZ or PROM1 for 24 h, serum-starved for 16 h, and treated with 5 ng/ml TGFβ for the indicated times. The expression levels and patterns of PROM1, p-SMAD2/3, total SMAD2/3 and SMAD7 were determined by immunofluorescence analysis (F) and/or immunoblotting (G). Scale bar = 20 µm. t test; *p < 0.05, **p < 0.01. All data are the mean ± S.E.M.
