Fig. 3

PG controls VE-PTP protein expression and phosphorylation of VE-cadherin at tyrosine 658. (a) Representative immunostaining for endothelial specific phosphatase VE-PTP and tyrosine phosphorylated VE-cadherin at position 658 (VE-cadherin, Tyr658), in WT and PG-KO cell monolayers; Stars denote intracellular staining in both cell lines, while arrowheads and arrows represent staining along the junctions in WT and PG-KO, respectively. DAPI was used to label nuclear DNA; N ≥ 3 (b) Corresponding Western blot analyses, where α-tubulin was used as a loading control. The bands originated from the same membrane and the vertical, continued line indicates excised irrelevant bands. Original blots are presented in Figure S4 p) provided in the Supplementary info file; N = 5 (c) Bar diagram depicting the protein expression of VE-PTP and VE-cadherin, Tyr658 in PG-KO cells, relative to the expression in WT. For VE-cadherin, Tyr658 the bar diagram represents the ratio between VE-cadherin, Tyr658 and total VE-cadherin. All data were normalized to the WT controls, (d) PCR analysis revealing the VE-PTP mRNA expression in WT and PG-KO. Equal loading was validated with B2M; N = 7 e) Bar diagram presenting the VE-PTP mRNA expression in PG-KO, relative to WT. Data are presented as mean ± SEM; unpaired t-test; * p ≤ 0,05; ** p ≤ 0,01.