(A) HEK293 cells were transfected with 5 ng pCMV-empty (-SIRT1, -S) or 5 ng pCMV-SIRT1 (+SIRT1, +S) in addition to 50 ng of pSG5-VDR and 250 ng of pTZ18U as carrier DNA. The cells were treated post-transfection with 10 nM 1,25D, 10 nM 1,25D+20 µM EX-527 (EX), or 10 nM 1,25D+25 µM Res. Quantitative PCR was performed on cell lysates and results are presented as a fold-induction after normalizing to human GAPDH. Error bars represent standard deviation. Results are the average of 15 biological replicates (n=15). (B) As in (A) except that cells were treated with ETOH, 10 nM 1,25D, 10 nM 1,25D+20 µM EX-527 (EX), or 10nM 1,25D+25 µM Res. Note that the group treated with EX-527 also received an additional dose of drug during time of transfection to inhibit endogenous SIRT1. Results shown are the average of nine biological replicates (n=9). P-values are noted in the figure panels.