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. 2016 Mar;44(3):352–355. doi: 10.1124/dmd.115.068437

Fig. 1.

Fig. 1.

Effect of PPAPDC2 overexpression on SQ1-mediated CAR activation in primary cultured rat hepatocytes. (A) Twenty-four hour–old primary cultures of rat hepatocytes were transiently transfected with a CAR-responsive reporter plasmid (CYP2B1-Luc) and either an empty expression plasmid (Empty) or an expression plasmid for rat (rPPAPDC2) or human PPAPDC2 (hPPAPDC2). Then, 24 hours after transfection, cultures were incubated in medium alone (UT) or containing 0.1 µM SQ1 or 100 µM PB. Each bar represents the mean ± S.E.M. of the mean normalized (firefly/Renilla) luciferase measurements from four independent experiments (n = 4). All values are normalized to the Empty/UT group. *P < 0.05; **P < 0.01 (significantly different from Empty/SQ1 group). (B) Rat hepatocytes were cotransfected with CYP2B1-Luc and either empty vector (Empty) or rPPAPDC2 and treated with medium alone (UT), 0.1 μM SQ1, 0.1 μM SQ1 and 30 μM pravastatin (Prava), or 100 μM PB. Forty-eight hours after drug treatment, transfected cultures were harvested for measurement of luciferase activity. Each bar represents the mean ± S.E.M. of mean normalized (firefly/Renilla) luciferase measurements from three independent experiments (n = 3). All values are normalized to the Empty/UT group. *P < 0.05 (significantly different from the Empty/SQ1 group). (C) Rat hepatocytes were transduced with empty adenoviral vector (Ad-375) or adenoviral vector expressing rPPAPDC2 (Ad5.CMV-PPAPDC2) and harvested after 48 hours for measurement of either rat PPAPDC2 or β-actin protein. The bar graph shows band intensity quantification of PPAPDC2 levels normalized to β-actin. (D) Rat hepatocytes were transduced with empty adenoviral vector (Ad-375) or adenoviral vector expressing rPPAPDC2 (Ad5.CMV-PPAPDC2), treated for 48 hours with medium alone (UT) or 0.1 μM SQ1, and harvested for measurement of CYP2B1 mRNA levels. Each bar represents the mean CYP2B1 levels ± S.E.M from four independent experiments normalized to the Ad-375/UT group (n = 4). *P < 0.05 (significantly different from Ad-375/SQ1 group).