Fig. 7
ERG drives thrombomodulin expression in vitro selectively in low shear stress conditions. a Representative image and quantification of ERG (green) and KLF2 (magenta) expression in HUVEC cultured under static conditions or exposed to low shear stress (LSS, 5 dynes per cm2) or high shear stress (HSS, 20 dynes per cm2) for 24 h by immunofluorescence; nuclei are identified by DAPI (blue). Scale bar 40 µm. Quantification represents the mean pixel intensity for ERG or KLF2 signal (arbitrary unit, A.U.) per cell (n = 3–4 images per condition). b Representative image of proximity ligation assay (PLA) for nuclear ERG-KLF2 interaction was performed on confluent HUVEC cultured under static conditions or exposed to low shear stress (LSS, 5 dynes per cm2) or high shear stress (HSS, 20 dynes per cm2) for 24 h; nuclei are identified by DAPI (blue). Quantification represents the PLA signal (mean number of dots per cell) and is expressed as a percentage compared to static condition (n = 3–4 images per condition). Scale bar, 40 μm. c qPCR analysis of ERG, KLF2 and TM mRNA expression in siCtrl or siERG-deficient HUVEC under static conditions or after 24 h under LSS or HSS (n = 4). d qPCR analysis of ERG, KLF2 and TM mRNA expression in siCtrl or siERG-deficient HDBEC under static conditions or after 24 h under LSS or HSS (n = 3 independent experiments). e Representative image and quantification of H3K27Ac expression (magenta) in siCtrl or siERG-deficient HUVEC cultured under static conditions or exposed to LSS or HSS for 24 h by immunofluorescence; nuclei are identified by DAPI (blue). Scale bar 40 µm. Quantification represents the mean pixel intensity for H3K27Ac signal (arbitrary unit, A.U.) per cell (n = 6 images per condition). All graphical data are mean ± s.e.m., *P < 0.05, **P < 0.01, ***P < 0.001, Student’s t-test for top panel and one-way Anova for bottom panel. Source data are provided as a Source Data file
