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. 2021 Jul 14;8:679518. doi: 10.3389/fmed.2021.679518

Figure 4.

Figure 4

Podocyte Myo9A expression and SNO-Myo9A are regulated by glucose and NO. (A) qPCR shows that Myo9A mRNA is not affected by mannitol, decreases ~50% in podocytes exposed to high glucose and addition of NO donor prevents Myo9A mRNA downregulation, mean ± SD, n = 4 independent experiments; Welch's ANOVA p < 0.02, unpaired t-test with Welch's correction: n.s. control vs. mannitol, p < 0.02 control vs. high glucose, p < 0.02 high glucose vs. high glucose + DETA. (B) Immunoblots show that Myo9A protein expression is not altered by mannitol, decreases ≥50% in podocytes exposed to high glucose and addition of NO donor prevents Myo9A downregulation. (C) BST shows SNO-Myo9A in control podocytes, SNO-Myo9A ~50% decrease in podocytes exposed to high glucose, addition of NO donor partially prevents Myo9A de-nitrosylation. Input shows total Myo9A loading, mean ± SD, n = 3–5 independent experiments, Brown-Forsythe ANOVA test, p = 0.022, unpaired t-test with Welch's correction non-significant (n.s.) control vs. mannitol, **p = 0.0046 control vs. high glucose, p = 0.0575 (n.s.) high glucose vs. high glucose + DETA. (D) Migration ‘wound' assay shows that podocyte migration is not affected by mannitol, whereas high glucose clearly reduces podocyte migration and addition of NO donor partially prevents this defect, mean ± SD, n = 4 independent experiments; Welch's ANOVA p < 0.0001, unpaired t-test with Welch's correction non-significant (n.s.) control vs. mannitol, p < 0.005 control vs. high glucose, p < 0.02 high glucose vs. high glucose + DETA. *p < 0.05, **p < 0.01, ****p < 0.0001.