CCA cells (HuCCT1, TFK-1, KKU-452, KKU-100, and QBC939) and human intrahepatic biliary epithelial cells (HIBEpiC) were purchased from ATCC. The DMEM medium (Invitrogen, USA) containing 10% fetal bovine serum (Gibco, USA) was used for cells. The culture conditions were 37°C and 5% CO2 in an incubator.
The cells were inoculated into 6-well plates according to the density of 2.5 × 105/well. sh-eIF5A#1 (5′-GCATTACGTAAGAATGGCTTT-3′), sh-eIF5A#2 (5′-GCATTCAAGATGGTTACCTTT-3′), sh-eIF5A#3 (5′-GCCATGTAAGATCGTCGAGAT-3′), shRNA-NC (5′-TTCTCCGAACGTGTCACGT-3′), pcDNA, and pcDNA-eIF5A were obtained from GenePharma (Shanghai, China). As previous studies [23 (link)], after overnight of culture, the serum-free medium was replaced, and then transfection was carried out by lipofectamine 3000 (Invitrogen). After 6 h of culture, medium was replaced; then the cells were cultured for 24 h. Following that, subsequent experiments were performed.
The cells were inoculated into 6-well plates according to the density of 2.5 × 105/well. sh-eIF5A#1 (5′-GCATTACGTAAGAATGGCTTT-3′), sh-eIF5A#2 (5′-GCATTCAAGATGGTTACCTTT-3′), sh-eIF5A#3 (5′-GCCATGTAAGATCGTCGAGAT-3′), shRNA-NC (5′-TTCTCCGAACGTGTCACGT-3′), pcDNA, and pcDNA-eIF5A were obtained from GenePharma (Shanghai, China). As previous studies [23 (link)], after overnight of culture, the serum-free medium was replaced, and then transfection was carried out by lipofectamine 3000 (Invitrogen). After 6 h of culture, medium was replaced; then the cells were cultured for 24 h. Following that, subsequent experiments were performed.
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