Skip to main content
. 2021 Oct 16;24(11):103296. doi: 10.1016/j.isci.2021.103296

Figure 1.

Figure 1

ATG5 shows opposite effects on cell growth under normal culture conditions and serum starvation conditions

(A) ATG5 was depleted or overexpressed in HeLa cells with two ATG5-specific siRNAs (left) or a Flag-tagged ATG5 construct (right), respectively. HeLa cells transfected with a control siRNA (left) or Flag control construct (right) were used as the corresponding controls. Cells were further cultured under normal culture conditions for the indicated durations, and the relative cell growth ability was examined by a cell counting kit 8 (CCK-8) assay.

(B) HeLa cells transfected as described in A were cultured under serum starvation conditions for the indicated durations. The relative cell growth ability was examined as described in (A).

(C) HeLa cells were transfected with a Flag control or a Flag-tagged ATG5 construct and cultured under normal conditions (left) or serum starvation conditions (right) for 6 h. Cells were then treated with or without 1 μM Baf A1 as indicated and were further cultured for different durations as indicated. The relative cell growth ability was examined by CCK-8 assays as described in (A).

(D) HeLa cells stably transfected with an ATG5-specific shRNA or a Flag-tagged ATG5 construct were generated, and a colony formation assay was performed with cells cultured under normal conditions (left upper) or serum starvation conditions (left lower) for 7 days. The colonies were photographed (left), and the relative quantification of the number of colonies is shown (middle and right).

(E) HeLa cells were transfected with a Flag control or a Flag-tagged ATG5K130R construct and cultured under normal conditions (left) or serum starvation conditions (right) for 6 h. Cells were then treated with or without 1 μM Baf A1 as indicated and were further cultured for different durations as indicated. The relative cell growth ability was examined by CCK-8 assays as described in (A). Statistics: all experiments were repeated more than 3 times. The bars and error bars indicate the mean ± s.d. values; n = 3 independent replicates. Two-tailed unpaired Student's t test was performed. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001; ns: nonsignificant.