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. 2020 Dec 14;11:6387. doi: 10.1038/s41467-020-19974-5

Fig. 6. G-1749 acts on pre-associated IRE1.

Fig. 6

a, b Western blots for indicated proteins of KMS-11 (a) and MDA-MB-231 (b) cell lysates after treatment of the cells for 4 h with indicated compounds. DMSO was used as a negative control (0.06%) and Thapsigargin (Tg, 100 nM) as a positive control. Before being loaded on a gel, lysates were crosslinked with disuccinimidyl suberate (DSS, 250 nM) for 1 h at room temperature, quenched with TRIS buffer (1 M) for 15 min at room temperature. One representative out of two experiments is shown. c Comparison of IRE1 and XBP1s protein concentration in, left, KMS-11 and MDA-MB-231 cell lines, compared to, right, MDA-MB-231 IRE1−/− after transfection with IRE1 kinase-dead (D688N) using a CMV promoter. d Kinase-dead (D688N IRE1) under the control of a CMV promoter was overexpressed in MDA-MB-231 IRE1 KO cells using a lipofectamine kit. Transfected cells were then treated with compounds for 4 h. Reported are western blots of indicated proteins after crosslinking with DSS. One representative out of two experiments is shown. Source data for all blots are provided as a Source Data file.