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. 2021 Aug 23;12:724141. doi: 10.3389/fphar.2021.724141

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Copyright © 2021 Shi, Hu, Wang, Ma, Tang, Tao, Qiu, Zhuang and Liu.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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3-MA decreases autophagic activity in cultured human peritoneal mesothelial cells. (A) Photomicrographs illustrate light microscope observation of HPMCs. (B) Cell lysates were subjected to immunoblot analysis with specific antibodies against LC3, Beclin-1 and GAPDH. (C, D) Expression levels of LC3II and Beclin-1 were quantified by densitometry and normalized with LC3I and GAPDH respectively. (E) Photomicrographs show immunofluorescence co-staining of Beclin-1 and DAPI. (F) The count of Beclin-1-positive cells. (G) Transmission electron microscopy images of autophagosome (Yellow arrows) in HPMCs following TGF-β1 (2 ng/ml) stimulation in the presence/absence of 3-MA. (H) Quantitation of the number of autophagic vacuoles per cell. Data are represented as the mean ± SEM. Means with different superscript letters are significantly different from one another (p < 0.05). Scale bars in (A) = 500 μm, (E) = 100 μm, (F) = 10 μm.