Polyplus in vivo jetpei

RIPK1 and RIPK3 were silenced in C₃H mice in vivo by transfecting with 6 μg of RIPK1 vs. negative control siRNA (Bioneer, Daejeon, Korea) using Polyplus in vivo-jetPEI (Polyplus-transfection, Illkirch, France) as a transfection reagent with a transfection efficiency of ~ 70% as previously described^{16 (link),17 (link)}.
See Experimental scheme (Fig. 5B).

SV-HUC-1, TCCSUP, 5637, BIU-87, T24, 293 T cells were purchased from the Chinese Academy of Science (Shanghai, China). Cells were cultured in RPMI-1640, DMEM or Ham’s F-12 medium (Procell Co., Ltd, China) and supplemented with 10% FBS (Biological Industries, Israel) at 37 °C in a 5% CO₂ humidified atmosphere. The TEAD4 was overexpressed in the BLCA cells by transfection with the plasmid (Genechem, Shanghai, China) in Polyplus Invivo-jetPEI (Polyplus, French) reagent according to the instructions recommended. To construct of TEAD4 stable knockdown cells lines, 5637 and T24 cells in logarithmic growth phase were transfected by lentivirus, following by culturing in 1640 medium with 10% FBS in a 6-well dish. Puromycin (2 μg/μl) was added for selection when the cell density reached 90%, and the stable colnoies will be amplified after 10–14 days. The overexpression/knockdown efficiency of TEAD4 was evaluated by qPCR and western blot.