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. 2021 Dec 13;4:1391. doi: 10.1038/s42003-021-02906-4

Fig. 4. D6 promotes L858R/T790M-EGFR degradation.

Fig. 4

a, b Immunoblotting analysis (a) of cell lysates derived from NCI-H1975 cells treated by CHX (50 μg/ml) in the presence or absence of D6 (2.5 μM); protein quantification is presented in (b), n = 3 independent experiments. P values were calculated by two-way ANOVA analysis. c EGFR protein levels were detected by immunoblotting in NCI-H1975 cells treated by D6 (2.5 μM) with or without MG132 (10 μM). d NCI-H1975 or A549 cells transfected with HA-Ub were then treated with or without D6 (2.5 μM) for 6 h. Cell lysates were immunoprecipitated with anti-HA beads and beads elution was further analyzed by immunoblotting. e, f Immunoblotting analysis (e) of cell lysates derived from HEK293 cells transfected with different EGFR mutants and then were subjected to CHX (50 μg/ml) chase assay with or without D6 (2.5 μM) incubation. Protein degradation rate was shown as curves in (f), n = 3 independent experiments. P values were calculated by two-way ANOVA analysis. g Immunoblotting analysis of the elution of anti-HA beads prepared from HEK293 cells transfected with HA-Ub and EGFR mutant with or without D6 (2.5 μM) exposure. Data are presented as mean ± SEM (b and f). Representative results were analyzed from at least three independent experiments.