siRNA and CRISPR/single guide RNA (sgRNA) treatment.

ECs were transfected with non-targeting control siRNA (Qiagen (1027280) or Thermo Fisher Scientific (AM4611)) or with siRNAs targeting Snail (Thermo Fisher Scientific; s13185), Slug (Thermo Fisher Scientific; s13127), ZEB1 (Thermo Fisher Scientific; s229971 or 4392420), MEF2D (Dharmacon; L-009884-00-0005), LEF1 (Dharmacon; L-015396-00-0005), PPAR-γ (Dharmacon; L-003436-00-0005) or PAK4 (Thermo Fisher Scientific; s20134 or 4390824) using RNAiMAX Lipofectamine (Invitrogen; 13778075) in serum-free Opti-MEM medium (Gibco; 31985-070) for 12 h, followed by incubation with serum-supplemented medium for 24 h. The guide RNA sequence (5′-GTGTTTGGGAAGAGGAAGAAG-3′) targeting PAK4 was subcloned into lentiCRISPRv2 puro vector (Addgene; 52961). ECs were transduced with the lentivirus expressing PAK4 sgRNA, followed by cell selection by incubation with medium containing 2 μg ml⁻¹ puromycin.