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

Figure 3.

Figure 3.

High glucose downregulates Bcl-2 expression via HK2 expression. HUVECs were transfected with HK2 adenovirus or control virus for 24 h. Following transfection, HUVECs were cultured in DMEM containing 5.5, 16.5 or 33 mM glucose for 72 h. (A) The relative mRNA expression level of Bcl-2 was determined by RT-qPCR in HUVECs. (B) HUVECs were co-transfected with pCF CREB (or pcDNA3.1) and pGL3-Bcl-2 (or pRL-TK-Renilla) or pCMV4-p65 (NF-κB) (or pCMV4) and pGL3-Bcl-2 (or pRL-TK-Renilla) for 48 h and luciferase activity was detected using the Dual-Luciferase reporter assay. The protein expression of CREB or NF-κB were increased in HUVECs following transfection with pCF CREB or pCMV4-p65 compared with pcDNA3.1 or pCMV4, respectively. (C) The protein expression levels of p-NFκB, NFκB, p-CREB and CREB were determined by western blot analysis in HUVECs. Data are presented as the mean ± standard error. *P<0.05 as indicated. HK2, hexokinase 2; HUVEC, human umbilical vein endothelial cell; DMEM, Dulbecco's modified Eagle's medium; NFκB, nuclear factor-κB; CREB, cyclic AMP response element binding protein; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; IB, immunoblotting.