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. 2023 Mar 6;19(8):2196–2216. doi: 10.1080/15548627.2023.2178876

Figure 4.

Figure 4.

CLU promotes mitochondrial fission by regulating DNM1L activity in oral CD44+ cells. (A-E) Western blot, confocal microscopy and mitochondrial morphology analyses. Western blot analysis for DNM1L phosphorylation status in CD44+ and CD44− cells, with (A) MOCK or CLU expression (left panel), cells expressing shNC or shCLU (right panel), (C) cells expressing shNC or shCLU and treated with vehicle or cisplatin (10 µM, 24 h) and (D) DNM1L KD cells expressing MOCK or CLU transfected with wild-type DNM1L or DNM1LS616A mutant. ACTB was used as a loading control. (Bi) Representative confocal images of the corresponding cells stained with CLU (red), and TOMM20 (green; mitochondria) (Ei) with showing mitochondrial morphology. (Bii, Biii, Eii) ImageJ-based quantification of mitochondrial branch length, calculated from images with multiple cells randomly taken for each sample of three different experiments, which was further categorized into fragmented (<2.5 µm), intermediate (2.5–4 µm), and elongated (>4 µm) according to the indicated parameter. Scale bars: 10 μm. Error bars: ±S.D. *p < 0.05, **p < 0.01, ***p < 0.001.