FIGURE 6.

Knockdown of ZEB2 expression in glomerular podocytes abrogates GH-dependent decrease in E-cadherin expression. A, immortalized human podocytes cultured under growth-permissive conditions were transduced with lentiviral constructs expressing either ZEB2 shRNA (shRNAs 1 and 2) or scrambled shRNA. Following transduction, cells were expanded, and aliquots were induced to differentiate for 14 days prior to harvesting of RNA for analysis. Steady state ZEB2 mRNA abundance was measured by RT-qPCR; GAPDH was used as an internal control. The results (n = 4–5) are depicted as mean ± S.E. The steady state abundance of the ZEB2 transcript is depicted relative to ZEB2 mRNA abundance in non-transduced naïve cells. B, immortalized human podocytes transduced with lentiviral constructs expressing either ZEB2 shRNA (shRNAs 1 and 2) or scrambled shRNA were differentiated for 14 days and then subjected to Western blot analysis for ZEB2 and tubulin as detailed under “Experimental Procedures.” Results shown are representative of two independent experiments. C, immortalized human podocytes transduced with lentiviral construct expressing either scrambled shRNA (left panel) or ZEB2 shRNA 2 (right panel) were differentiated, exposed to hGH (500 ng/ml), and subjected to Western blot analysis for E-cadherin and tubulin as detailed under “Experimental Procedures.” Densitometric analysis of the E-cadherin band, normalized for respective tubulin expression, is depicted relative to expression prior to exposure to GH. Error bars indicate mean ± S.E.; n = 4. *, p < 0.05 (Kruskal-Wallis test) compared with expression prior to exposure to GH.