Figure 8.

Fgf2 deficiency diminishes the paracrine effect of renal tubular cells on fibroblasts. (A) Subconfluent isolated primary proximal tubular cells from WT (Fgf2 WT PT) and fgf2 KO mice (fgf2 KO PT) were exposed to 5 ng/ml TGFB1 in serum-free DMEM for 3 days. Control cells were kept in serum-free medium without TGFB1. Both cell lysates and culture media were collected for immunoblots of cellular and secreted FGF2 (n = 3 experiments). (B-E) Subconfluent NRK-49F fibroblasts were incubated with CM from control WT primary proximal tubular cells (Fgf2 WT PT control-CM), TGFB1-treated WT primary proximal tubular cells (Fgf2 WT PT TGFB1-CM), control KO primary proximal tubular cells (fgf2 KO PT control-CM), or TGFB1-treated KO primary proximal tubular cells (fgf2 KO PT TGFB1-CM) for 2 days (n = 8 experiments). (B) Cell morphology was monitored by phase contrast microscopy. Scale bar: 100 µm. (C) Cell number counting and cellular protein measurement. (D) Immunoblots of COL1A1, FN1 and ACTA2. (E) Densitometry of COL1A1, FN1 and ACTA2 proteins. Data in (C and E) are presented as mean ± SD. Two-way ANOVA with multiple comparisons was used for statistics.