Journal of Aging Research / 2020 / Article / Fig 5

Research Article

Insulin-Like Growth Factor I Prevents Cellular Aging via Activation of Mitophagy

Figure 5

IGF-1 dose dependently upregulated Nrf2 and Sirt3 expression (a) and Nrf2/Sirt3 pathway mediated IGF-1-induced mitophagy activation (b, c). A, P20 SMC was treated with IGF-1 and Nrf2, and Sirt3 and β-actin levels were quantified by immunoblotting. (b, c) P20 SMC was transfected with 5 nM Nrf2 siRNA or Sirt3 siRNA or scrambled siRNA (control), and after 48 hours, cells were exposed to IGF-1, bafilomycin (Baf), or combination of IGF-1 + Baf. Mitophagy was detected by live cell staining with Mtphagy + Lyso dyes and quantifying a number of mitophagic vacuoles/cell (b) or by assessing mitochondrial LC3 II/I ratio (c). vs. untreated cells; # for SMC with IGF-1 vs. no IGF-1.

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