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. 2016 Apr 1;7(5):680–688. doi: 10.1111/jdi.12505

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© 2016 The Authors. Journal of Diabetes Investigation published by Asian Association for the Study of Diabetes (AASD) and John Wiley & Sons Australia, Ltd

This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial‐NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

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Effect of vitamin D (VD) and vitamin D receptor (VDR) small interfering ribonucleic acid (siRNA) transfection on lipopolysaccharide (LPS)‐mediated podocyte apoptosis. (a) Pre‐incubation with VD presented to protect podocytes from excessive apoptosis induced by LPS. (b) VDR‐siRNA transfection in podocytes resulted in further elevated apoptosis after LPS challenge. When the nuclear factor‐κB (NF‐κB) pathway was blocked by pyrrolidine dithiocarbamate (PDTC), the apoptosis rate showed an extreme reduction. (a,b) The results detected through flow cytometry by using an Annexin V‐FITC/PI staining kit. Q1, Q2, Q3 and Q4 in the figures represented dead cells, late apoptotic cells, early apoptotic cells and viable cells. Cell proportions of Q2 together with Q3 were calculated. (c,d) The results found through Hoechst 33 258 staining, which showed similar outcomes of (a) and (b), respectively. The apoptosis rate of cells was evaluated by scoring the cell numbers showing a pycnotic nucleus or fragmented nucleus. *P < 0.05. NCsi, negative control‐siRNA; VDRsi, VDR‐siRNA.