Figure 6.

Stat3 inactivation in tubular cells increases inflammatory cytokine transcriptional response. (A–C) Quantification of (A) Ccl5, (B) Cxcl10, and (C) Ccl2 mRNA abundance in mIMCD3 cells stably expressing shRNAs targeting STAT3 (sh1 and sh2) or a scramble control shRNA (scr) treated with LPS or vehicle for 24 hours. Bars are mean±SEM of eight independent experiments. (D) Representative Western blot and (E) quantification of three independent experiments of STAT3 phosphorylation and RELA expression in whole-cell lysates from mIMCD3 cells expressing the indicated shRNAs, 24 hours after LPS or vehicle treatment. (F) Representative Western blot and quantification of four independent experiments of (G) STAT3 and (H) RELA expression in nuclear extracts from mIMCD3 cells expressing the indicated shRNAs, 24 hours after LPS or vehicle treatment. Whole-cell lysates (WCL) served as a positive control for tubulin labeling. (A–C, E, G, and H). Dots represent independent experiments. Two-way paired ANOVA followed by Tukey–Kramer test, *P<0.05, **P<0.01, ***P<0.001.