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. Author manuscript; available in PMC: 2020 Sep 23.
Published in final edited form as: Dev Cell. 2019 Aug 1;50(6):690–703.e6. doi: 10.1016/j.devcel.2019.07.010

Figure 3. Autophagy-Dependent Cancer Cells Can Undergo Selection to Circumvent Inactivation of an Autophagy Regulator.

Figure 3.

(A and B) Incucyte quantification of mCherry+/GFP cell count after transfection with gRNAs targeting GFP and the indicated genes. Data are represented as mean ± SEM for technical replicates (N = 2–3). Graphs are representative of 2–3 individual experiments. Statistical analysis: two-way ANOVA.

(C and D) Western blot analysis in clonal isolates of mCherry+/GFP cells weeks after treatment with gATG7. Data are representative of 2–3 experiments.

(E–H) Incucyte quantification of mCherry+ cell count of WT or ATG7−/− clones in (E and F) nutrient replete conditions or (G and H) starved conditions in EBSS. Data are represented as mean ± SEM for technical replicates (N = 2–3) and graphs are representative of 2–3 individual experiments. Statistical analysis: two-way ANOVA.

(I and J) Normalized CGS for WT or ATG7−/− clones calculated from Incucyte quantification of mCherry+/GFP cell count normalized to essential and non-essential genes after transfection with gRNAs targeting GFP and the indicated genes. Data are represented as mean ± SEM for technical replicates (N = 2–3) and graphs are representative of 2 individual experiments. Statistical analysis: two-way ANOVA. *p % 0.05, **p % 0.01, ***p % 0.001, **** p % 0.0001. Statistical significance indicated for the last time point for time course graphs. See also Figure S5.