FIGURE 5.

NF‐κB inhibitor abrogates HK2 nonmetabolic function‐induced proliferation, migration, and invasion of pancreatic ductal adenocarcinoma (PDAC) cells in vitro. (D, E) Immunoblot analyses were performed with the indicated antibodies. (A) PANC‐1 and AsPC‐1 with or without HK2 D209/D657A expression were cultured in the presence or absence of JSH‐23 (10 μM) for the indicated time. Cell proliferation was examined using a Cell Counting Kit‐8 (CCK‐8) assay. Data are presented as the means ± SD from three independent experiments (n = 3). *p < 0.05; **p < 0.01; ***p < 0.001. (B) PANC‐1 and AsPC‐1 cells with or without HK2 D209/D657A expression were cultured in the presence or absence of JSH‐23 (10 μM) for 2 weeks before counting colony numbers. Data are presented as the means ± SD from three independent experiments (n = 3). **p < 0.01; ***p < 0.001. (C) The migration and invasion of the indicated cells in the presence or absence of JSH‐23 (10 μM) were examined by transwell assay. The membrane was photographed using a digital camera mounted onto a microscope. Scale bars, 50 μm. Data are presented as mean ± S.D. **p < 0.01; ***p < 0.001. (D) PANC‐1 and AsPC‐1 cells with or without HK2 D209/D657A expression were cultured in the presence or absence of JSH‐23 (10 μM) for 24 h. (E) PANC‐1 and AsPC‐1 cells with depleted HK2 and reconstituted expression of WT rHK2, rHK2 D209/D657A, or KD were cultured with or without hypoxia for 24 h.