Figure 6.

CTGF silencing reduces the COL1A1 production induced by TGF-β1. CTGF silencing was evaluated for TGF-β1- and FFA-induced CTGF and COL1A1 deposition. Spheroids were maintained and treated as shown in Figure 1. Blue bars present the mRNA level data, red bars present results of IHC analysis and green bars, data acquired using ELISA. (A) Silencing of CTGF was confirmed at the transcriptional level 10 days after seeding and transfection (n = 3). (B) CTGF silencing also reduced baseline pro-collagen I α1 and CTGF released in the medium in the last 3 days of culture (n = 3). (C,D) CTGF siRNA-transfection caused a decrease in TGF-β1-induced CTGF and COL1A1 production (n = 3). TGF-β1-induced COL1A1 production was significantly lower in the spheroids with silenced CTGF. (E,F) CTGF siRNA-transfection also caused a decrease in FFA-induced CTGF and COL1A1 production, although the difference was not significant. Every dot in the graphs represents data from 1 experiment and those in the red bars present an average of at least 10 analyzed sections of spheroids per experiment. Data are shown as mean ± SEM. Statistical analysis—Student’s t-test; * p < 0.05; ** p < 0.01.