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. 2019 Feb 19;8(3):12. doi: 10.1038/s41389-019-0124-4

Fig. 3. Fbw7 regulates XBP1s ubiquitination.

Fig. 3

a Deficient of Fbw7 leads to increased exogenous XBP1 expression. HCT116 Fbw7−/− cells or HCT116 control cells were transfected with pLenti 6.3 XBP1. Cycloheximide (CHX) at 80 μg/ml was used to treat HCT116 cells at the indicated time points. Western blotting was carried out with the indicated antibodies. b Deficient of Fbw7 leads to increased endogenous XBP1 expression. HCT116 Fbw7−/− cells or HCT116 control cells were treated with tunicamycin (TM) which is well-known ER stress inducer. c HCT116 Fbw7−/− cells were transfected with Flag-Fbw7 WT (wild type), Mt (mutant), XBP1s, and/or His-tagged ubiquitin (His-Ub) or vector control as indicated. Cells were then lysed in a buffer containing 6 M urea. WCL whole cell lysate. Ubiquitin-conjugated proteins were captured with nickel-agarose beads and subjected to immunoblot analysis with the indicated antibodies. d WT of Fbw7−/− HCT116 cells were transfected with His-tagged ubiquitin and/or XBP1s expressing vector. Ubiquitin-conjugated proteins were captured with nickel-agarose beads and subjected to immunoblot analysis with antibodies against XBP1 and actin