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. 2023 Mar 22;8(6):e158429. doi: 10.1172/jci.insight.158429

Figure 6. Lipogenesis drives OPA1 expression and mitochondrial fusion in CSCs.

Figure 6

(A) A549 tumor spheres were treated with the indicated doses of C75 for 12 hours and detected for OPA1 expression by qPCR. Mean ± SEM from 10 independent experiments. (B and C) A549 tumor spheres were treated with an increasing dose of C75 for 24 hours and detected for OPA1 protein with immunoblots. Representative and mean ± SEM from 5 independent experiments. (D) A549 tumor spheres were transfected with FASN siRNA or the control and detected for OPA1 expressions by qPCR after 12 hours. Mean ± SEM from 6 independent experiments. (E) A549 tumor spheres were transfected using a second FASN siRNA with or without the reintroduction of FASN by overexpression and were analyzed for OPA1 expression. Mean ± SEM from 8 independent experiments. (F and G) A549 tumor spheres with or without C75 treatment (50 μM) were detected for mitochondria morphology and areas. Scale bars: 500 nm. Representative and mean ± SEM from 5 independent experiments. Red arrows point to tubulated and elongated mitochondria. (H) A549 tumor spheres with or without C75 treatment (50 μM) were detected for mitochondrial OCR using Seahorse assay. Mean ± SEM from 5 independent experiments. **P < 0.01, ***P < 0.001 with ANOVA plus Tukey’s method (A–C and E) and paired t test (D and G).