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. 2022 Sep 27;11(19):3021. doi: 10.3390/cells11193021

Figure 5.

Figure 5

Metformin failed to activate Nrf2 in presence of Compound C(CC), an AMPK inhibitor. SRA-hLECs (A) and mLECs (B) were culture in 60 mm culture plate overnight, next day cells were treated with an AMPK inhibitor Compound C (CC) and followed by Metformin for 24 h as indicated. Total proteins were isolated and processed for SDS-PAGE immunoblotting with phospho-AMPKα (SRA-hLECs: (A,a) and mLECs: (B,a)), AMPKα (SRA-hLECs: (A,b) and mLECs: (B,b)) and Nrf2 (SRA-hLECs: (A,c) and mLECs: (B,c)) antibodies for protein expression analyses as shown in Figure. Below the protein bands, densitometric analysis of the protein band value normalized with corresponding tubulin and were presented as histograms. Figure (A) (SRA-hLECs) and (B) (mLECs) show increased phosphorylated form of AMPKα and Nrf2 (second lane in each figure) in response to Metformin treatment. However, Metformin failed to activate the AMPK and Nrf2 in presence of CC, an AMPK inhibitor (last lanes in each figure). Histograms are presented as mean ± S.D. values derived from three independent experiments, ** p < 0.001.