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. 2022 Feb 16;13(2):156. doi: 10.1038/s41419-022-04602-5

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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. 1 — A Whole-cell lysates of control, shCHCHD2, shCHCHD10, shCHCHD2/CHCHD10 HeLa cells were analyzed for indicated protein expression by immunoblot. β-Actin was used as a loading control. B Mitochondrial morphology in control, CHCHD2 knockdown, CHCHD10 knockdown, and CHCHD2/CHCHD10 knockdown HeLa cells immunostained for Hsp60 (Green) was visualized by confocal microscope. C Mitochondrial morphology described in B was counted according to the criteria detailed in Experimental Procedures (mean ± s.d. of n = 3 independent biological samples; two-way ANOVA with Tukey’s multiple comparisons correction, N.S. not significant). D Comparison of mitochondrial fusion and fission between control and shCHCHD2, shCHCHD10, shCHCHD2/CHCHD10 HeLa cells. Ten photoactivated mitochondria labeled with mito-PA-GFP were tracked by time-lapse microscopy for 20 min, and the number of mitochondrial fission and fusion events within 20 min was counted. Three independent cells were analyzed, the number of mitochondrial fission and mitochondrial fusions within 20 min were counted. Bars represent means ± S.D. of three independent experiments. Statistical significance analysis was used by two-way ANOVA with Sidak’s multiple comparisons test, *P < 0.05, **P < 0.01, ***P < 0.001, N.S. not significant. E Mitochondrial ultrastructure in control, shCHCHD2, shCHCHD10, shCHCHD2/CHCHD10 HeLa cells was analyzed by transmission electron microscope (TEM). F The relative number of mitochondria with abnormal cristae (ratio of abnormal mitochondria to total mitochondria) in control, shCHCHD2, shCHCHD10, shCHCHD2/CHCHD10 was counted. 100 mitochondria were measured from 20 cells. Statistical significance was assessed by one-way ANOVA with Dunnett’s multiple comparisons test; error bars represent means ± SD of three independent experiments; *P < 0.05,**P < 0.01,***P < 0.001, N.S., not significant.