Research Paper Volume 13, Issue 11 pp 14892—14909

Figure 5. CDK9 inhibitor reduces inflammatory responses and phenotype switch of VSMCs exposed to ox-LDL by suppressing NF-κB pathway. (A–B) VSMCs were treated with the Bay (10μM) for 1 h, and then stimulated with ox-LDL (50μg/mL) for 24 h. The expressions of α-SMA, Vimentin, and PCNA were detected by western blot ((n = 3; ^##P < 0.01, compared to control; ^*P < 0.05, ^**P < 0.01, compared to ox-LDL). (C–D) transfected with siRNA against CDK9 and then incubated with ox-LDL (50 μg/mL) for 1 h. The IκB-α protein level in cell lysate was detected by western blot; and the nuclear fraction was isolated and the nuclear level of NF-κB p65 was measured by western blot (n = 3; ^##P < 0.01, compared to control; ^*P < 0.05, ^**P < 0.01 compared to ox-LDL). (E) VSMCs were pretreated with LDC000067 (2.5 or 5 μM) for 1 h, and then stimulated with ox-LDL (50 μg/mL) for 1 h. Nuclear translocation of NF-κB p65 was measured by immunofluorescence staining (scale bar = 10 μm). (F–G) NF-κB-RE-EGFP reporter VSMCs were transfected with siRNA against CDK9 and then stimulated with ox-LDL (50 μg/mL) for 6 h. NF-κB activity is shown as mean fluorescence intensity (MFI) in flow cytometry histogram (n = 3; ^##P < 0.01, compared to control; ^**P < 0.01, compared to ox-LDL).