Rcor1

Short hairpin RNAs (shRNAs) against FOS, GATA2, MAFK, TEAD4, TFAP2C, NCOR2, RCOR1 and ZNF362 were purchased from Sigma-Aldrich (Supplementary Table S1). A total of 7.3 × 10⁶ HEK293T cells were transfected with 2.5 μg of the pLKO construct with a specific shRNA, 1.7 μg of Δ8.9 and 0.8 μg of VSVG using 15 μl of a GenJet transfection reagent (SignaGen) to generate lentiviruses expressing a specific shRNA. After 24 h, the medium was replaced with the TSC medium. After cultivating at 37°C for 24 h, the viral supernatant was collected, filtered with a syringe filter (0.45 μm) and used to infect TSCs. To knock down individual TFs, 2.5 × 10⁵ TSCs were infected with pLKO-shRNA viral particles in a well of a 12-well plate. Infected cells were incubated overnight, and the medium was replaced with fresh TSC medium supplemented with puromycin to select out the infected cells. To determine the KD efficiency of each TF and profile global gene expression, we harvested the cells after 3 days of selection to get complete KD of an individual TF.