Figure 7.

The COL-PGE₂ matrix favors renal tubular epithelial cell proliferation and cytoprotection in vitro

(A) Immunofluorescence staining of Ki-67 (green) in primary renal tubular epithelial cells (pRTECs) cultured on plates coated with 2.0 μM COL-PGE₂, PGE₂/COL, or COL matrix or uncoated plates. Scale bars, 100 μm.

(B) Expression levels of apoptosis-related genes were evaluated by qPCR.

(C) Immunofluorescence staining of Yap (red) and Sox9 (green) in pRTEC cultured in noncoated, COL, PGE₂/COL, or COL-PGE₂ at 48 h. Scale bars, 50 μm.

(D) Quantification of the Yap⁺ area and Sox9⁺ cells in (C).

(E) Immunoblot analysis of the Yap, p-Yap, p-Lats1, Areg, Survivin, and Sox9 protein in pRTEC cultured on noncoated, COL, PGE₂/COL, or COL-PGE₂ matrix at 48 h. β-tubulin was used as loading control.

(F) Immunofluorescence staining of Yap (red) and Sox9 (green) in pRTEC cultured in plates coated with COL or COL-PGE₂ and pRTECs with YAP knockdown cultured on plates at 48 h. Scale bars, 50 μm.

(G) Quantification of the Yap⁺ area and Sox9⁺ cells in (F).

(H) Western blot analysis of the Yap, p-Yap, p-Lats1, Areg, Survivin, and Sox9 protein in pRTEC cultured on COL- or COL-PGE₂-coated plates and pRTEC with Yap knockdown cultured on plates at 48 h. β-tubulin was used as loading control. One-way repeated measures ANOVA with Tukey post hoc tests (B and D) was used for statistical analysis. Data are expressed as mean ± SD; n = 3. ∗p < 0.05 versus COL; ^#p < 0.05 versus PGE₂/COL. One-way repeated measures ANOVA with Tukey post hoc tests (G) was used for statistical analysis. Data are expressed as mean ± SD; n = 3. ∗p < 0.05 versus the non-targeting shRNA (sh Ctrl) cultured in COL; ^#p < 0.05 versus the shRNA targeting YAP (sh-Yap) cultured in COL-PGE₂.