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

Figure 1.

Figure 1.

Cisplatin promotes mitophagy in oral CD44+ cells. (A-D) Oral cancer cells (FaDu, left panel; and CAL-33, right panel), the derived orospheres and MACS-sorted CD44− and CD44+ cells were analyzed for expression of (A, B) autophagic protein LC3-II, ATG5, and SQSTM1, (C, D) mitochondrial-, nuclear-, endoplasmic reticulum (ER)-, lysosome-, and endosome-related proteins through western blot. ACTB was used as a loading control to normalize band intensities. (E-F) Western blot analysis for mitochondrial proteins in CD44+ cells and CD44− cells of FaDu (left panel) and CAL-33 (right panel) in response to cisplatin treatment for dose-dependent (1, 5, and 10 µM) and time-course (6, 12 and 24 h) studies. ACTB was used as a loading control to normalize band intensities. (Gi-ii) Immunofluorescence-based quantification of the number of mitophagosomes in CSC compared to their non-stem counterparts exposed to 10 µM of cisplatin for 24 h using MitoTracker Green (MTG) and LysoTracker Red (LTR). The graph shows Pearson’s coefficient values ±S.D., calculated from images with multiple cells randomly taken for each sample of three different experiments. Scale bars: 10 μm. *p < 0.05, **p < 0.01.