Fig. 2.

IRE1α-XBP1 signaling during exposure of primary mouse hepatocytes to EFV or 8-OHEFV. Representative gel images and sXBP1/uXBP1 densitometry quantification ratios of XBP1 mRNA splicing from semiquantitative reverse transcriptase PCR in (A) male mouse primary hepatocytes incubated with 10, 20, 30, 40, or 50 μM EFV for 4 hours (mean ± S.D, n = 5), and (B) male mouse primary hepatocytes with 50 μM EFV for 5, 15, 30 minutes, 1, 2, or 4 hours (mean ± S.D., n = 4). (C) Representative images for Western blotting of p-IRE1α, total IRE1α, and β-Actin and densitometry quantification ratios of p-IRE1α/total IRE1α for male mouse primary hepatocytes treated with 10, 20, 30, 40, 50 μM EFV or 50 μM 8-OH EFV for 2 hours (mean ± S.D., n = 3). Ratios of sXBP1/uXBP1 and p-IRE1α/total IRE1α densitometry were log-transformed and the statistical significance of TM, EFV, or 8-OHEFV treatments from DMSO (0.1%) control was determined using an unpaired t test on the transformed data. For (B) treatments were compared with the DMSO control from the same time point. Two-tailed P values were generated for all analyses (*P < 0.05; **P < 0.01; ***P < 0.001).