Fig. 4: KRAS regulates DUSP6 stability.
a Western blots showing changes in p-ERK1/2, DUSP4, and DUSP6 levels in 293 T cells stably expressing HA-KRASG12V treated with trametinib or ulixertinib for 16 h. The bar graph includes quantification of three independent experiments. Data was presented as mean ± SEM. P-values were calculated using one-way ANOVA followed by Dunnet’s multiple comparisons test, HA-KRAS vs control (p = 0.0031), HA-KRAS vs 2 μM ulixertinib (p = 0.0474), HA-KRAS vs 0.5 μM trametinib (p = 0.0468), HA-KRAS vs 1 μM trametinib (p = 0.0045). Western blots showing serial changes in endogenous DUSP6 protein levels in 293 T cells stably expressing an empty vector or HA-KRASG12V (b) or in Pa01c cells stably expressing a scramble sequence or an shRNA against KRAS (c), after treatment with 10 μg/mL cycloheximide (CHX) for the indicated durations. d Western blots, and quantification of serial endogenous DUSP6 protein levels in Pa01c cells stably expressing a scramble sequence or an shRNA against KRAS (c), after treatment with 2 μM of ulixertinib for the indicated durations. e Western blots, and quantification of serial endogenous DUSP6 in Pa01c cells pre-treated with DMSO or MRTX1133 (0.5 μM) for 1 h followed by 2 μM of ulixertinib at the indicated durations. f Western blots, and quantification of serial endogenous DUSP6 in MIA Paca-2 cells pre-treated with DMSO or AMG-510 (0.5 μM) for 1 h, followed by 2 μM of ulixertinib for the indicated durations. For (b–f), half-lives (t1/2) of DUSP6 were calculated by measuring the DUSP6 band intensities, normalizing to t0 and performing one-phase exponential decay analysis, as shown in the graph below. a was conducted three times, and (b–f) were conducted two times, and one set of data for each was shown. Source data are provided in Source Data file.
