Knockdown of RUNX1, TSP1 and TGFβRII

RUNX1, TSP1 TGFβRII knockdown was achieved using lentiviral shRNA vectors from the Mission TRC genome-wide shRNA collections purchased from Sigma-Aldrich Corporation with the following catalog numbers; Scrambled shRNA#: SHC016, RUNX1#1: TRCN0000338428, RUNX1#2: TRCN0000338427, TSP1#1: TRCN0000226402, TSP1#2: TRCN0000219072, TGFβRII#1: TRCN0000000831 and TGFβRII#2: TRCN0000000834. Lentiviral supernatants were generated using the calcium phosphate method as described^{118 (link)}. Cancer cells were incubated with lentivirus-containing media with polybrene (8 µg/ml) and incubated for 72 h at 37 °C with 5% CO₂ followed by 1 µg/ml of Puromycin (Wisent Inc., 450-162-XL) selection for 15 days.