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. 2022 Feb 24;41:76. doi: 10.1186/s13046-022-02262-z

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© The Author(s) 2022

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — Drp1 overexpression regulates autophagy in ESCC cells. (A) RNA sequencing was performed on ESCC cells as indicated to screen differentially expressed mRNAs. Signaling pathway were plotted according to enriched gene ratio. (B) Heat map analysis of RNA sequencing for autophagy associated gene expression in ESCC cells as indicated. (C) Western blot analyses were performed in ESCC cells as indicated with antibodies specific for the proteins shown. (D) Representative transmission electron microscopy images of double-membrane autophagosomes in ESCC cells as indicated. Scale bar: 0.5 μm. (E) Representative images of fluorescent LC3B puncta (green) in ESCC cells as indicated. Scale bar: 10 μm. (F) Representative immunohistochemical (IHC) staining images of LC3B in xenograft tumors developed from KYSE-30 cells as indicated. Scale bar: 100 μm. (F) Colony-forming potential were detected in KYSE-30 cells as indicated. Scale bar: 10 mm. (G) Representative immunohistochemical (IHC) staining images of LC3B in paired ESCC tissues (n = 61). Scale bar: 100 μm. (H and I) Cell proliferation was evaluated by colony formation EdU incorporation assay in KYSE-30 cells as indicated. Scale bar: 200 μm. The data shown are the mean ± SEM from three separate experiments. Drp1, expression vector encoding Drp1; EV, empty vector. P value from t tests. *, P < 0.05; **, P < 0.01; ***, P < 0.001