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. 2019 May 24;18(1):621–629. doi: 10.3892/etm.2019.7609

Figure 4.

Figure 4.

High glucose inhibits HK2 transcription of HK2 via PPARγ downregulation in HUVECs. (A) HUVECs were cultured in DMEM containing 5.5, 16.5 or 33 mM glucose for 72 h and the relative mRNA expression level of HK2 was determined by RT-qPCR. (B) HUVECs were cultured in DMEM containing 5.5, 16.5 or 33 mM glucose for 72 h followed by treatment with MG132 (50 µM) for 8 h. Following 8-h treatment, the protein expression level of HK2 was determined by western blot analysis. (C) HUVECs were co-transfected with pSV SPORT PPARγ (or pSV-SPORT) and pGL3-HK2 (or pRL-TK-Renilla) into cells for 48 h and luciferase activity was detected using the Dual-Luciferase assay. HUVECs were cultured in DMEM containing 5.5, 16.5 or 33 mM glucose for 72 h. (D) The relative mRNA expression level of PPARγ was determined by RT-qPCR in HUVECs. (E) The protein expression level of PPARγ was determined by western blot analysis in HUVECs. (F) HUVECs were transfected with the Flag tagged PPARγ for 24 h. Following transfection, HUVECs were cultured in medium containing 5.5, 16.5 or 33 mM glucose for 72 h. The protein expression level of Flag was determined by western blot analysis in HUVECs. Data are presented as the mean ± standard error. *P<0.05 as indicated. HK2, hexokinase 2; PPARγ, peroxisome proliferator-activated receptor γ; HUVEC, human umbilical vein endothelial cell; DMEM, Dulbecco's modified Eagle's medium; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; MG132, cell-permeable proteasome inhibitor; IB, immunoblotting.