Figure 3 | Scientific Reports

Figure 3

From: Astragaloside IV inhibits glucose-induced epithelial-mesenchymal transition of podocytes through autophagy enhancement via the SIRT–NF-κB p65 axis

Figure 3

In vitro AS-IV effects on SIRT1 and NF-kB p65 expression in hyperglycaemia-triggered podocyte EMT. (A–F) Podocytes were exposed to either a SIRT1 inhibitor EX527 or SIRT1 activator SRT1720 after a 1 hour exposure to hyperglycaemic conditions, and were subsequently incubated with or without AS-IV (100 μM) for 48 hours. (A,E) mRNA expression levels of SIRT1and p65 were quantified using real-time PCR. (B) The deacetylase activity of SIRT1 was detected with a SIRT1 activity assay. (C,D,F) The protein levels of SIRT1 and AC-p65 were quantified using Western blotting. The molecular weight of the proteins: SIRT1, 110 kDa; AC-p65, 65 kDa; p65, 65 kDa. Data is presented as mean ± SD. n = 3. *Compared with normal glucose cohort, or AS-IV cohort, or high glucose plus AS-IV cohort, or high glucose plus SRT1720 cohort, P < 0.05; #compared with high glucose plus AS-IV cohort, P < 0.05. (G) Podocytes were pretreated with high glucose for 1 hour, and then incubated with PDTC or AS-IV for 48 hours. The levels of TGF-β, nephrin, E-cadherin, N-cadherin and α-SMA were quantified with Western blotting. Data is presented as mean ± SD. n = 3. *Compared with normal glucose cohort, or high glucose plus AS-IV cohort, or high glucose plus PDTC cohort, P < 0.05.

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