Fig. 6

mGEC exposed to diabetic serum secrete factors that cause podocyte cell death. a Percentage podocyte cell death (AnnexinV-FITC and PI positive cells) 24 h after co-culture with mGEC in transwell inserts; without or with HG, or HG in the presence of mitoTEMPO (5μg/ml), or CS, or DS, or DS + mTEMPO for 24 h (n = 4). b Percentage apoptotic podocytes after supernatant transfer (SN) from mGECs; NG-SN, or HG-SN, or HG + mTEMPO-SN, or CS-SN, or DS-SN, or DS + mTEMPO-SN. MitoTEMPO dose used was 5 μg/ml (n = 3). c Representative immunofluorescence images detecting cleaved caspase 3 (red), and F-Actin (phalloidin; green) and nuclei (DAPI; blue) in podocytes cultured with NG-SN, or HG-SN, or CS-SN, or DS-SN, or DS + mTEMPO-SN for 24 h. Scale = 50 μm. Cultures were performed in 10% FCS. Bars in a and b represent averaged independent experiments ± s.e.m.; *P < 0.05, **P < 0.01 versus NG or CS controls, +P < 0.05, ++P < 0.01 versus HG, #P < 0.05 versus DS