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. 2020 Jan 27;219(2):e201904048. doi: 10.1083/jcb.201904048

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© 2020 Grey et al.

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Figure S5. — Expression and XBP1 splicing activity of IRE1 constructs. Supports Fig. 6. (A) IRE1α protein was assayed by immunoblot (top), spliced and unspliced XBP1 transcript was assayed by PCR and gel electrophoresis (middle), and spliced XBP1 transcript was assayed by qPCR (bottom) for HEK293 or HEK293/IRE1α^KO cells treated with DMSO, Tg, or tunicamycin (Tm; n = 3). (B) IRE1 protein expression was assayed by immunoblot (top), and spliced XBP1 transcript was assayed by qPCR (bottom) for HEK293/IRE1α^KO cells transfected with indicated expression vector and treated with DMSO, Tg, or tunicamycin (n = 3). (C) XBP1 splicing luciferase reporter activity for IRE1α constructs transfected in HEK293/IRE1α^KO cells. Activity is plotted relative to the untreated (no Tg), mock-transfected cells. Bars represent mean of two independent experiments. Bars and error bars represent mean values ± SEM; significance is indicated by asterisks (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001).