Primary MAoECs (passage 3; PELOBiotech GmbH, Planneg, Germany) and primary HAoECs (passages 2, 3; Promocell, Heidelberg, Germany) were cultured using endothelial cell complete growth medium (Promocell) containing gentamicin (0.05 mg ml−1; ThermoFisher).
To grow the cells under different shear stresses, MAoECs were cultured in collagen-coated perfusion chambers (µ-Slides VI0.4, ibidi GmbH, Martinsried, Germany) and exposed to high shear stress (10 dyne cm−2) or low shear stress (5 dyne cm−2) for 48 h generated by the perfusion with EC complete medium (ibidi Pump System, ibidi GmbH) containing 5-ethynyl-2’-deoxyuridine (EdU, 10 µM final conc., Click-iT® EdU Alexa Fluor® 488 Imaging Kit, Life Technologies) and TSBs (TSBs in vivo ready) or control LNAs (both 50 nM final conc., Exiqon).
MAoECs were transfected with antisense oligonucleotides to block the interaction between miR-103 and lncWDR59 (TSB; 50 nM, miRCURY LNATM microRNA Target Site Blockers; Exiqon) or scrambled controls. To inhibit lncWDR59 function, murine and human ECs were transfected with lncWDR59 GapmeRs (50 nM, LNATM GapmeRs; Exiqon) that strongly induce the degradation of the lncWDR59 transcript in the nucleus of the ECs.
To grow the cells under different shear stresses, MAoECs were cultured in collagen-coated perfusion chambers (µ-Slides VI0.4, ibidi GmbH, Martinsried, Germany) and exposed to high shear stress (10 dyne cm−2) or low shear stress (5 dyne cm−2) for 48 h generated by the perfusion with EC complete medium (ibidi Pump System, ibidi GmbH) containing 5-ethynyl-2’-deoxyuridine (EdU, 10 µM final conc., Click-iT® EdU Alexa Fluor® 488 Imaging Kit, Life Technologies) and TSBs (TSBs in vivo ready) or control LNAs (both 50 nM final conc., Exiqon).
MAoECs were transfected with antisense oligonucleotides to block the interaction between miR-103 and lncWDR59 (TSB; 50 nM, miRCURY LNATM microRNA Target Site Blockers; Exiqon) or scrambled controls. To inhibit lncWDR59 function, murine and human ECs were transfected with lncWDR59 GapmeRs (50 nM, LNATM GapmeRs; Exiqon) that strongly induce the degradation of the lncWDR59 transcript in the nucleus of the ECs.