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. 2015 Apr 8;26(12):2963–2977. doi: 10.1681/ASN.2014050528

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Copyright © 2015 by the American Society of Nephrology

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Figure 7. — TxNIP knockdown in cultured human podocytes exposed to high glucose inhibits the increased mitochondrial O₂⁻, membrane potential, and apoptosis. Knockdown of TxNIP attenuates mitochondrial O₂⁻, and membrane potential and apoptosis in cultured human podocytes. Podocytes were transfected with 50 nM TxNIP-specific siRNA (siTxNIP) or universal negative control siRNA (scrambled, scr) for 24 hours and then incubated in 5 mM normal glucose (NG) or 25 mM HG for 48 hours. (A and B) Mitochondrial superoxide formation and mitochondrial membrane potential were assessed by confocal microscopy using MitoSox and JC-1 dyes, respectively (n=3), with the quantification illustrated as graphs in the bottom panel. (C) Lactate concentration in the cell culture media (n=6). Apoptosis in podocytes treated with NG/HG and scr/siTxNIP was assessed with (D) TUNEL assay and (E) cleaved caspase-3 protein expression in three independent experiments. Results are mean±SEM. *P<0.05; **P<0.01; and ***P<0.001 versus scr NG; ^#P<0.05 and ^###P<0.001 versus scr 48-hour HG.

Figure 7. — TxNIP knockdown in cultured human podocytes exposed to high glucose inhibits the increased mitochondrial O₂⁻, membrane potential, and apoptosis. Knockdown of TxNIP attenuates mitochondrial O₂⁻, and membrane potential and apoptosis in cultured human podocytes. Podocytes were transfected with 50 nM TxNIP-specific siRNA (siTxNIP) or universal negative control siRNA (scrambled, scr) for 24 hours and then incubated in 5 mM normal glucose (NG) or 25 mM HG for 48 hours. (A and B) Mitochondrial superoxide formation and mitochondrial membrane potential were assessed by confocal microscopy using MitoSox and JC-1 dyes, respectively (n=3), with the quantification illustrated as graphs in the bottom panel. (C) Lactate concentration in the cell culture media (n=6). Apoptosis in podocytes treated with NG/HG and scr/siTxNIP was assessed with (D) TUNEL assay and (E) cleaved caspase-3 protein expression in three independent experiments. Results are mean±SEM. *P<0.05; **P<0.01; and ***P<0.001 versus scr NG; ^#P<0.05 and ^###P<0.001 versus scr 48-hour HG.