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. 2022 May 30;19(1):751–763. doi: 10.1080/15476286.2022.2079889

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© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — Hypoxia-induced autophagy of CC cell is mediated by DARS-AS1/ATG5/ATG3 signalling. A HeLa cell viability was detected at different times after hypoxia treatment in the presence of 3-MA via CCK-8. B Impacts of DARS-AS1 up-regulation or down-regulation on the protein levels of p62 and LC3 in HeLa cells treated under normoxia or hypoxia treatment were investigated via western blot. C Fluorescence images of GFP-mRFP-LC3 stably expressed in DARS-AS1-silenced HeLa cells under hypoxia or DARS-AS1-overexpressed HeLa cells under normoxia were observed. The puncta visible due to GFP (green) and RFP (red) revealed the formation of autophagosomes. D-E RT-qPCR and western blot measured the mRNA and protein levels of autophagy-related genes in DARS-AS1-silenced cells under hypoxia. F Western blot assay detected the protein levels of p62 and LC3II in hypoxia-treated HeLa cells after transfection of sh-DARS-AS1 and ATG5/3 on under hypoxia. Experimental procedures were repeated independently for three times. *P < 0.05, **P < 0.01.