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. 2020 Jul 24;11:1114. doi: 10.3389/fphar.2020.01114

Figure 3.

Figure 3

Metformin activates AMPK/SIRT1-mediated autophagy in cultured chondrocytes. Representative Western blots (A) and densitometric quantification (B) of p-AMPKα, AMPKα, SIRT1, p62, and LC3B in the chondrocytes treated with different concentration of metformin in the presence of IL-1β. Expressions of target protein were normalized to GAPDH and shown as means ± SD. *P < 0.05 compared to control group, #P < 0.05 compared to the cells treated with IL-1β (n=5). Representative Western blots (C) and densitometric quantification (D) of p-AMPKα, AMPKα, SIRT1, Collagen II, MMP13 treated with AMPK inhibitor Compound C or SIRT1 inhibitor EX527 in IL-1β-stimulated (10ng/ml) chondrocytes with or without metformin (1mM) for 24h. Expressions of target protein were normalized to GAPDH and shown as means ± SD. *P < 0.05 compared to control group or as indicated (n=5). AMPK, 5’ adenosine monophosphate-activated protein kinase; SIRT1, silent mating type information regulation 2 homolog1; p-AMPK, Phospho- 5’ adenosine monophosphate-activated protein kinase; p62, Sequestosome-1; LC3B, Microtubule-associated proteins 1A/1B light chain 3B; MMP13, Matrix Metallopeptidase 13; EX527, Selisistat; IL, Interleukin; GAPDH, Glyceraldehyde 3-phosphate dehydrogenase; SD, standard deviation.