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. 2019 Jun 24;116(28):14154–14163. doi: 10.1073/pnas.1813580116

Fig. 2.

Fig. 2.

Caspase 12 contributes to podocyte ER stress-mediated apoptosis at the early stage of NS. (A) Primary podocytes from Tg-WT, Tg-C321R, and Lamb2+/− mice at P27 were stained with FITC-Annexin V and PI, and analyzed by flow cytometry. The values in the lower right quadrant of the representative histograms indicate the percentage of podocytes undergoing early apoptosis (Annexin V+/PI− cells). The percentage of early apoptotic podocytes was expressed as the mean ± SD from 3 independent experiments. ***P < 0.001 by ANOVA. PE, phycoerythrin. (B and C) Primary podocyte lysates from Tg-WT, Tg-C321R, and Lamb2+/− mice at P27 were analyzed by WB with the indicated antibodies. Arrows indicate cleaved forms of the proteins of interest. Quantification of cleaved caspase 12 (B), as well as cleaved caspase 3 (C), was normalized to β-actin. (D) Glomeruli were isolated from age-matched Tg-WT, Tg-C321R, and Lamb2+/− mice at P27. Cleaved caspase 12, cleaved caspase 3, and β-actin were analyzed by WB in isolated glomeruli from the indicated genotypes. Quantification of cleaved caspase 12 and cleaved caspase 3 was normalized to β-actin in isolated glomerular lysates. Arrows indicate cleaved (active) caspases 12 and 3. Mean ± SD of 5 independent experiments. *P < 0.05, **P < 0.01 by ANOVA.