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. 2024 Jan 30;300(3):105695. doi: 10.1016/j.jbc.2024.105695

Figure 4.

Figure 4

Impact of AMPKα expression on BHLHE40-expressing EC cells. Immunoblotting analysis of EC cells transfected with an siRNA against AMPKα1/2 (siAMPKα) and cultured for 24 h in DMEM with 10% FBS and 1 mM sodium pyruvate without glucose. HEC1 (A) and Ishikawa (B) cells. A and B, values under panels indicate relative expression levels of p- AMPKα/AMPKα, p-ACC/ACC, p-PDHA1/PDHA1, p-LDHA/LDHA, AMPKα/ACTB and LDHA/ACTB. A and B, data are representative of at least three biological replicates. PDH activity (C and E) and LDH activity (D and F) were measured in HEC1 (C and D) and Ishikawa (E and F) cells after culturing for 24 h in DMEM with 10% FBS and 1 mM sodium pyruvate without glucose. (C–F) Data are from three technical replicates. The experiments were biologically replicated three times and representative data are shown. Extracellular flux analysis of HEC-1 (G and H) and Ishikawa (I and J) cells. Real-time OCRs (G and I) and ECARs (H and J) were measured upon treatment with the indicated inhibitors or glucose. G–J, data are from three technical replicates. The experiments were biologically replicated twice and representative data are shown. C–F, unpaired two-sided Student’s t test or the Mann–Whitney U test was used. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. LtCtrl, LtControl; LtE40, LtBHLHE40; shCtrl, shControl; shE40, shBHLHE40.