Skip to main content
. 2023 Aug 9;12(8):1586. doi: 10.3390/antiox12081586

Figure 1.

Figure 1

Mutation of the AMPK-dependent phospho-sites to alanine stabilizes Nrf2 in Keap1−/− cells in a βTrCP2-dependent manner. (A,B). Keap1−/− MEFs were transfected with EGFP-MYC tagged WT- or TM-Nrf2 expression plasmids for 48 h and then exposed to cycloheximide (50 μM) for different periods of time. Cell lysates were subjected to immunoblot analysis for MYC or β-Actin. A representative blot (A) and quantification analysis (B) of n = 9 independent experiments (relative to signal at t = 0) are depicted. One-phase exponential decay curves were fitted to the data by finding the least sum of squares using GraphPad Prism. The curves were significantly different, with a p value = 0.0108. (C,D) Keap1−/− MEFs were transfected with Fbxw11-specific siRNA and with EGFP-MYC tagged WT- or TM-Nrf2 expression plasmids for 48 h, and then exposed to cycloheximide (50 μM) for different periods of time. Cell lysates were subjected to immunoblot analysis for MYC or β-Actin. A representative blot (C) and quantification analysis (D) of n = 5 independent experiments (relative to signal at t = 0) are depicted. One-phase exponential decay curves were fitted to the data by finding the least sum of squares using GraphPad Prism. There was one curve for all data sets with a p value = 0.4852. The dotted lines in (B,D) represent the confidence bands with a 95% confidence level. Data are presented as means  ±  SEM.