Figure 4

CAGE CRISPR-Cas9 cell lines show decreased autophagic flux and growth rates. (a) Cell lysates from the indicated cancer cell line were subjected to immunoblot. CAGE CRISPR-Cas9 cell lines were transfected with the indicated construct for 48 h, followed by immunoblot. The uncropped blots are shown in Supplementary Materials. (b) Cellular proliferation was determined by trypan blue exclusion assays. Significance determined by one-way ANOVA. ***, p < 0.001. (c) Colony forming potential of each cancer cell line was determined as described. Significance determined by one-way ANOVA. ***, p < 0.001. Cells were seed at 400 cells/dish. (d) Each indicated cell line was treated with various concentrations of osimertinib for 24 h. MTT assays were performed. Significance determined by one-way ANOVA. ***, p < 0.001. (e) Each CAGE CRISPR-Cas9 cell line was subjected to tumor spheroid forming potential assays. Immunoblot was also performed. Data are presented as mean ± SEM. Significance determined by one-way ANOVA. **, p < 0.01. The uncropped blots are shown in Supplementary Materials. (f) Invasion potential of the indicated cancer cell line was determined. Data are presented as mean ± SEM. Immunoblot was also performed. Significance determined by one-way ANOVA. ***, p < 0.001. The uncropped blots are shown in Supplementary Materials.