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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 4. — IRE1β enzymatic activity is not required to suppress stress-induced XBP1 splicing. (A) Schematic of IRE1β constructs. All constructs have C-terminal Myc tag. Expression of constructs in HEK293T cells was assayed by immunoblot with anti-Myc antibody. (B and C) XBP1 splicing activity was measured using a luciferase reporter in HEK293T cells cotransfected with indicated IRE1β expression plasmids and stimulated with either Tg or SubAB. Luciferase activity is plotted (B) as the fold change relative to mock-transfected, control-treated cells at baseline or (C) as the stress-induced fold change relative to control for a given IRE1β construct (n = 3). 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).