Figure 4. Newly Acquired Autophagy Independence Causes Resistance to Pharmacological Autophagy Inhibition.

(A) Incucyte mCherry⁺ cell count/mm² in BT549 WT or ATG7^−/− clones treated with CQ (50 mM ) normalized corresponding untreated wells. The data are represented as mean ± SEM for technical replicates (N = 3) and are representative of 4 individual experiments. Statistical analysis: two-way ANOVA.

(B) The Incucyte mCherry⁺ cell count/mm² 72 h after treatment with the indicated dose of CQ in BT549 WT or ATG7^−/− clones normalized to corresponding untreated samples. Data are represented as mean ± SEM for technical replicates (N = 3) and are representative of 3 individual experiments. Statistical analysis: two-way ANOVA.

(C) Normalized Caspase3/7 CellEvent green count after treatment with CQ (50 mM) for BT549 WT and ATG7^−/− clones. The data are represented as mean ± SEM for technical replicates (N = 3) and are representative of 2 individual experiments. Statistical analysis: two-way ANOVA.

(D) Representative Incucyte images of Caspase 3/7 CellEvent green 48 h after treatment with 100 mM CQ in BT549 WT and ATG7^−/− clones. The scale bars represent 0–200 mM.

(E) Normalized Caspase3/7 CellEvent green count 72 h after treatment with CQ in BT549 WT and ATG7^−/− clones. Data are represented as mean ± SEM for technical replicates (N = 3) and representative of 2 individual experiments. Statistical analysis: two-way ANOVA.

(F) Mean tumor volume as a percentage of pre-treatment baseline, following initiation of vehicle or HCQ-treatment in mice bearing H292 WT or ATG7^−/− xenograft tumors. N = 5–7 mice per group. Data are represented as mean ± SEM. 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.