Piperlongumine disrupts the interaction of HK2 and VDAC1 and induces apoptosis. A, HCC827 (left) and H1975 (right) cells were treated with piperlongumine for 24 h, co-immunoprecipitation was conducted to detect the interaction between HK2 and VDAC1. B, HCC827 cells were treated with piperlongumine for 24 h, co-immunoprecipitation and western blot were performed to identify the phosphorylation of HK2. C, HK2-WT or HK2-T473D was transfected into HCC827 cells, the co-immunoprecipitation and western blot were conducted to determine the interaction between HK2 and VDAC1 as indicated. D, HCC827 (left) and H1975 (right) cells were treated with piperlongumine for 24 h, cytosolic fractions and mitochondrial fractions were isolated, western blot was conducted to detect the target proteins as indicated. E, HCC827 (left) and H1975 (right) cells were treated with piperlongumine for 24 h, the whole cell extract was subjected to western blot analysis as indicated. F, HCC827 (left) and H1975 (right) cells were treated with piperlongumine for 24 h, the flow cytometry was conducted for apoptosis analysis. PL, Piperlongumine. G and H, HCC827 (G) and H1975 (H) stable cells were treated with piperlongumine for 48 h, western blot and MTS assay were conducted. Asterisk, significant (*p<0.05, **p<0.01) difference between groups as indicated. ns, not statistically significant.