MCF-7 wild-type and MCF-7 AJAP1:EGFP cells were maintained in DMEM (GIBCO) supplemented with 10% FBS (GIBCO) and 2 mM L-Glutamine (GIBCO) at 37°C, and 5% CO2. Pre-screened HUVECs (C-12205, Lot: 0061801, 1071101.1, 2012601.1, PromoCell, Heidelberg, Germany) were, if not stated otherwise, maintained in EBM (CC-3121, Lonza, Basel, Switzerland) supplemented with hydrocortisone, bovine brain extract, epidermal growth factor (all from Lonza), and 10% FBS (GIBCO) at 37°C, and 5% CO2.
Experiments with HUVECs were approved by: Declaration of Helsinki; German Federal Data Protection Act; Human Tissue Act; and Convention for the protection of Human Rights and Dignity of the Human Being with regard to the Application of Biology and Medicine: Convention on Human Rights and Biomedicine.
12.5 µM small Interfering RNAs (siRNAs) (Sigma Aldrich, St. Louis, MO USA) were transfected using peqFECT (Peqlab, Erlangen, Germany) for AJAP1 down-regulation during the spheroid-based sprouting assay, or using Lipofectamine RNAiMAX (Thermo Fisher, Waltham, MA, USA) or Interferin (Peqlab, Erlangen, Germany) in the wound healing assay according to manufacturer's instructions. siAJAP1-1: SASI_Hs01_00244567; siAJAP1-2: SASI_Hs02_00352284; siAJAP1-3: SASI_Hs01_00244565. As a control, a scrRNA (SIC002-10NMOL) was used.
Experiments with HUVECs were approved by: Declaration of Helsinki; German Federal Data Protection Act; Human Tissue Act; and Convention for the protection of Human Rights and Dignity of the Human Being with regard to the Application of Biology and Medicine: Convention on Human Rights and Biomedicine.
12.5 µM small Interfering RNAs (siRNAs) (Sigma Aldrich, St. Louis, MO USA) were transfected using peqFECT (Peqlab, Erlangen, Germany) for AJAP1 down-regulation during the spheroid-based sprouting assay, or using Lipofectamine RNAiMAX (Thermo Fisher, Waltham, MA, USA) or Interferin (Peqlab, Erlangen, Germany) in the wound healing assay according to manufacturer's instructions. siAJAP1-1: SASI_Hs01_00244567; siAJAP1-2: SASI_Hs02_00352284; siAJAP1-3: SASI_Hs01_00244565. As a control, a scrRNA (SIC002-10NMOL) was used.