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. 2022 Dec 9;7:388. doi: 10.1038/s41392-022-01253-y

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — Knocking down CRL4^CUL4A/DDB1 leads to mitochondrial fragmentation in cisplatin-resistant OCCs. a (Left panel) Representative images of mitochondrial morphology in (upper panel) A2780CP and (lower panel) COC1/DDP cells with CRL4^CUL4A/DDB1 knockdown stained with MitoTracker Green. Images were captured by a laser confocal microscope. Results represent the mean from three independent experiments, measured in quadruplicate. (Right panel) Quantification analyses of the mitochondrial footprint in CRL4-knockdown (upper panel) A2780CP and (lower panel) COC1/DDP cells. The significance of differences was calculated using Student’s t-test (***p < 0.001). b DRP1 expression level in CRL4-knockdown cells detected by western blot, GAPDH serves as an internal loading control. Protein levels were quantified with Image J software and normalized to the control group (NT). c Immunoblot detection of ACC1, p-ACC1, CUL4A, DDB1, DRP1, p-DRP1^Ser616, p-DRP1^Ser637, AMPK-α1, and p-AMPK-α1 in (left panel) A2780CP and (right panel) COC1/DDP cells with CRL4^CUL4A/DDB1 knockdown. β-actin serves as an internal loading control. d Immunoblot detection of MFF, p-MFF, CUL4A, DDB1, AMPK-α1, and p-AMPK-α1 in (left panel) A2780CP and (right panel) COC1/DDP cells with CRL4^CUL4A/DDB1 knockdown. e Immunoblotting analysis of DRP1 and MFF in the cytosolic (Cyto) and mitochondrial (Mito) fractions of A2780CP cells with CRL4^CUL4A/DDB1 knockdown. f Immunoblot detection of DRP1, AMPK-α1, and VDAC1 in (upper panel) input or (lower panel) immunoprecipitated samples in the NT control or CRL4^CUL4A/DDB1 knockdown (left panel) A2780CP and (right panel) COC1/DDP cells. β-actin serves as a loading control