Podocyte injury and activation of the mTOR-ULK1 pathway.

<p>(A-B).Western blot analysis of cellular proteins confirmed that the most significant upregulated expression of mTOR signaling pathway-associated proteins was observed in 2 h puromycin amino nucleotide (PAN)-treated differentiated mouse podocyte cells (MPCs) compared to that in the control group and the other time points of treated cells, demonstrating the obvious activation of the mTOR-ULK pathway, *<i>P</i><0.05 vs Control. (C, D, F).At this same time point, the expression of synaptopodin decreased and the filamentous expression (red fluorescence) was barely visible; therefore, we selected 2 h as the modeling time. PAN-treated podocytes showed an increase in ULK1 phosphorylation, a decrease in LC3 II expression, and a decrease in cellular GFP-LC3 granular expression (green fluorescence), indicating the inhibition of autophagy.(E).Compared to GFP-LC3 transfected cells, there was no granulation in GFP transfected cells. Magnification = ×1200.