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. 2020 Sep 15;11:561306. doi: 10.3389/fphar.2020.561306

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Copyright © 2020 Xiao, Chen, Jiang, Chen, Chen, Zhu, Zhang, Huang, Qin, Zhang, Yi and Luo

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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Ubc9 promotes the formation and the autophagy-positioning of PI3K-III complexes Adv-Ubc9 was transfected in NRCMs for 48 h, then maintained in hypoxic condition for 6 h. (A, B) Endogenous PI3K-III complexes in NRCMs. Vps34 was IP, and followed by IB for Beclin1, Vps15, ATG14, and UVRAG. (C) Fixed cells were immunostained with antibodies of Vps34 (red) and LC3 (green), and confocal microscopy for colocalization to percentage of Vps34 and LC3, n = 4 per group. (D) Fixed cells were immune-stained with antibodies of Vps34 (red) and Lamp1 (green), confocal microscopy for colocalization to percentage of Lamp1 and Vps34. Magnifications of the boxed area are shown in the right; arrows point to colocalization sites of Vps34 with Lamp1, n = 4 per group. Data were analyzed by one-way ANOVA, followed by a Bonferroni post-hoc test.