FIG. 6.

Drug-induced changes in UPR gene expression after ThG or TuM in MEFs. Gstp1/p2^+/+ and Gstp1/p2^−/− MEF cells were treated with ThG (200 nM) or TuM (500 μg/ml) for certain times as indicated. Proteins were separated by SDS-PAGE and UPR proteins, including BiP, IRE1, ATF6, phosphor-eIF2α, eIF2α, ATF4, PDI, and CHOP evaluated by immunoblots. Even loading of proteins was confirmed by probing with anti-actin antibodies. Goat anti-mouse, goat anti-rabbit, and donkey anti-goat fluorescent secondary antibodies were used and immunoblots imaged on a two-channel IR fluorescent Odyssey CLx imaging system. Data are representative blots from three independent experiments (A, B). The activation of XBP1 was evaluated by normal PCR for unsliced and sliced XBP1(C). Relative gene expression levels were quantified by real-time PCR. Bars represent the mean (±SD) from three independent experiments (D, E). *Represents significant difference between control and treatment by UPR inducer.