Figure 9

5Aza and TSA elicit increases in PPARα/γ expression, and PPAR antagonists block 5Aza and TSA-mediated increase in cubilin expression. A-B, qPCR analysis of PPARα and γ mRNA expression in RNA isolated from NRK cells cultured for 60 h with 3 medium changes each containing the indicated doses of 5Aza. C and D, qPCR analysis of PPARα and γ mRNA expression in RNA isolated from NRK cells cultured for 24 h with the indicated doses of TSA. E and F, qPCR analysis of cubilin and megalin was performed on RNA from NRK cells treated for 36 h with 2 medium changes each containing 5 μM of 5Aza and then a 24 h treatment with 5Aza-free medium containing PPAR antagonists (each at 10 μM). G and H, qPCR analysis of cubilin and megalin mRNA in RNA isolated from NRK cells cultured for 24 h with TSA alone (100 nM) or TSA (100 nM) plus PPAR antagonists (each at 10 μM). Cubilin, megalin and PPARα and γ mRNA levels were normalized to GAPDH. Asterisks indicate that differences between mRNA levels in vehicle control and drug treatment groups were significant at p<0.05. Pound signs indicate that differences between mRNA levels in drug treatment groups alone and drug treatment with PPAR antagonists were significant at p<0.05.