Fig. 6: PEAK1 mediates extracellular matrix (ECM)-regulated VEGFR2 mRNA transcription in human ECs.

a–c Bar graphs represent relative mRNA levels vs. HPRT1 measured by quantitative RT-PCR (qPCR) and normalized against siCtrl. Mean ± SEM; n = 3. a, b Cells were cultured on collagen I-coated plates. a HUVECs or HMVECs were treated with siRNAs. P value, vs. siCtrl in each target mRNA. b HUVECs were treated with siRNAs and transfected with or without non-targeted GFP-PEAK1 (Rescue) or GFP only; c HUVECs were attached to plastic dishes coated with either collagen I (Colla), fibronectin (Fibro), or left uncoated (Un) and treated with different siRNAs. d HUVECs treated as in (c) were transfected with the plasmid of luciferase driven by VEGFR2 promoter. Bar graph represents relative luciferase activity measured by illuminometer and normalized against siCtrl + Uncoated group. Mean ± SEM; n = 4. e Confocal images of siCtrl and siPEAK1 treated HUVECs stained with GATA2 or GTF2I antibodies (green) along with DAPI, as a nuclear stain (blue). Cells were cultured on collagen I coated coverglass. Scale bar = 10 μm. f WB of HUVECs attached to different coated plates and treated with siRNAs. g Bar graph shows relative protein level vs. α-tubulin from HUVECs treated as in (f). Relative densitometry from three independent WB were analyzed by Image J. Mean ± SEM; n = 3. All data are representative of at least three independent experiments. ***P < 0.001; **P < 0.01; *P < 0.05; N.S., not significant