Fig. 6

Caspase-10 represses ACLY promoted malignant phenotype a, b A549 control (control), caspase-10 knockdown (CASP10kd), caspase-10/ACLY double knockdown (CASP10kd/ACLYkd), caspase-10/GCN5 double knockdown (CASP10kd/GCN5kd), and caspase-10/ACLY/GCN5 triple knockdown (CASP10kd/ACLYkd/GCN5kd) cells were subjected to glucose starvation for 48 h. The cells were treated with sodium acetate (5 mM), palmitic acid (0.2 mM) or both for the last 24 h of starvation period as indicated. a In vitro invasion, and (b) migration potential of the cells was then measured. Error bars are means ± SD of three biological replicates. Statistical analyses were done using one-way ANOVA (Tukey’s post hoc test). ***P < 0.001. c A549 control, A549 CASP10kd, A549 CASP10kd/ACLYkd, A549 CASP10kd/GCN5kd, and A549 CASP10kd/ACLYkd/GCN5kd cells were seeded in soft agar and maintained in glucose starvation (5 mM) conditions, and treated with sodium acetate (5 mM), palmitic acid (0.2 mM) or both as indicated. Scale bars: 200μm. Representative images of colonies are shown. d A549 control (control), A549 CASP10kd, A549 CASP10kd/ACLYkd, and A549 CASP10kd/GCN5kd cells were injected subcutaneously into nude mice. Post-one week of injection, mice were treated with metformin (5 mg/ml in drinking water). Tumor volume was measured on the indicated days. The data shown are representative of three biological replicates (n = 5 mice/independent experiment). Error bars represent mean ± SD from five individual mice. Statistical analyses were done using one-way ANOVA (Tukey’s post hoc test). ****P < 0.0001. e At the end of 30 days, tumors were excised and weighed. The data shown are representative of three biological replicates (n = 5 mice/independent experiment). Error bars represent mean ± SD from five individual mice. Statistical analyses were done using one-way ANOVA (Tukey’s post hoc test). *P < 0.05; ***P < 0.001. f Immunofluorescence staining for Ki67 was performed on tumor sections (panel d above). Results shown is the representative of staining in five individual biological samples performed in triplicates. Statistical analyses were done using one-way ANOVA (Tukey’s post hoc test). *P < 0.05; **P < 0.01; ***P < 0.001. Scale bar: 10 μm. g At the end of 30 days, lysates of tumors (panel d above) were analyzed by immunoblotting. The data shown are representative of three independent experiments. Source data are provided as a Source Data file