The human colorectal cancer cell line SW480 and HT-29 (both from epithelial colorectal adenocarcinoma) were purchased from the American Type Culture Collection (Rockville, MD). The SW480/Vector, SW480/NNMT-1, SW480/NNMT-2 and HT-29/NC, HT-29/NNMT shRNA 1# and HT-29/NNMT shRNA 2 # cells were constructed by the method in our previous study [15 (link)]. SW480 cells were cultured in RPMI-1640 medium and HT-29 cells were cultured in McCoy's 5A medium. All media were supplemented with 10% fetal bovine serum, 100 U/mL of penicillin and 100 μg/mL of streptomycin, and cells were maintained at 37°C in a humidified 5% CO2 incubator.
The following antibodies were obtained from Cell Signaling Technology (Beverly, Massachusetts, USA): anti-ASK1, anti-phospho-ASK1 (Thr 845), anti-caspase-3, anti-cleaved caspase-3, anti-caspase-9, anti-cleaved caspase-9, anti-p38, anti-phospho-p38, anti-GAPDH. The mouse anti-NNMT monoclonal antibody 1E7 was prepared through the hybridoma technique as previously described [19 (link)]. Goat anti-mouse and goat anti-rabbit HRP-conjugated antibodies were obtained from Zhongshan Goldenbridge Biotechnology Co. (Beijing, China).
The following antibodies were obtained from Cell Signaling Technology (Beverly, Massachusetts, USA): anti-ASK1, anti-phospho-ASK1 (Thr 845), anti-caspase-3, anti-cleaved caspase-3, anti-caspase-9, anti-cleaved caspase-9, anti-p38, anti-phospho-p38, anti-GAPDH. The mouse anti-NNMT monoclonal antibody 1E7 was prepared through the hybridoma technique as previously described [19 (link)]. Goat anti-mouse and goat anti-rabbit HRP-conjugated antibodies were obtained from Zhongshan Goldenbridge Biotechnology Co. (Beijing, China).
Free full text: Click here
