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. 2011 Dec 23;287(9):6266–6274. doi: 10.1074/jbc.M111.331264

FIGURE 1.

FIGURE 1.

Hsp90 inhibition induced IRE1α activation and degradation in INS-1 cells. A, INS-1 cells were treated with 1 μg/ml GA or 1 μm TM for 30 min and immunoblotted for p-IRE1α and total IRE1α after immunoprecipitation with IRE1α antibodies. β-Actin immunoblot analysis was performed on total extracts as a loading control. B, INS-1 cells were treated with 1 μg/ml GA for 30 or 180 min and with DTT for 30 min. Unfolded protein response proteins from total extracts were analyzed by immunoblotting as indicated. NT, non-treated. C, XBP-1 pro-RNA splicing was detected by RT-PCR from INS-1 cells treated with GA for 0.5, 1, or 2 h; 1 μm TM or 1 μm TG for 2 or 4 h; and DMSO for 2 h as indicated. D, INS-1 cells were cultured with fresh media containing 1 μg/ml GA every 30 min for 2 h, and immunoblot for p-IRE1α and XBP-1 protein as indicated. The bottom panel includes XBP-1 pro-RNA splicing detected by RT-PCR.