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. 2020 Jul 15;133(14):jcs241356. doi: 10.1242/jcs.241356

Fig. 2.

Fig. 2.

BRSK2 and BRSK1 inhibit NRF2-mediated transcription. (A) hQR41-luciferase assay in HEK293T cells following expression of control (hcRED, pGUS), positive control (MAFG, KEAP1), BRSK2 splice variants (BRSK2iso4, BRISK2 iso3) or mouse Brsk2. Cells were treated with vehicle control or CDDO-me (100 nM, 12 h). (B,D) BRSK2 overexpression inhibits endogenous NRF2 target gene (HMOX1, GCLM and SLC7A11) induction in HEK293T cells (representative of n=3). (E) Western blot of HEK293T cells 24 h after transfection with the indicated expression plasmid. Cells were treated with CDDO-me 12 h before lysis. (F) hQR41-luciferase reporter assay in HEK293T cells expressing the indicated plasmid combinations for 24 h. (G) hQR41-luciferase reporter assay in HEK293T cells expressing the indicated plasmids. Cells were treated with CDDO-me 12 h before lysis. (H) hQR41 reporter assay in wild-type MEFs and KEAP1−/− MEFs expressing the indicated plasmids. (I) HEK293T cells were transiently transfected with the indicated firefly transcriptional reporter, Renilla luciferase, hcRED, BRSK2 or BRSK1. Two-way ANOVA with multiple comparison was performed comparing BRSK2 or BRSK1 to hcRED control. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.