Reagents and Cells

Doxycyline hyclate (D9891) was from Sigma-Aldrich (USA). Rabbit monoclonal anti-neuropilin-1 (ab81321) was from Abcam (UK). Rabbit polyclonal anti–extracellular signal-regulated kinase (ERK1/2) (M-5670) and mouse monoclonal anti-FLAG (F1804) were from Sigma-Aldrich. Rabbit polyclonal anti-Tet-repressor antibody (TET01) was from MoBiTec (Germany). The secondary horseradish peroxidase–conjugated rabbit anti-mouse IgG (P0260) and swine anti–rabbit IgG (P0217) were from DAKO (Belgium). PC3 human prostate adenocarcinoma cells were cultured in F-12 Kaighn's medium (Invitrogen, Belgium) supplemented with 7% fetal bovine serum (FBS) (Lonza, Belgium). MDA-MB-231 human breast adenocarcinoma cells were cultured in DMEM (Lonza) supplemented with 10% fetal bovine serum. Human skin fibroblasts, HSMC (human smooth muscle cells), HUVEC (human umbilical vein endothelial cells), HT1080 (human fibrosarcoma cells) and MCF7 (human breast adenocarcinoma cells) were cultured in DMEM supplemented with 10% FBS, MO59J (human malignant glioblastoma cells) were cultured in DMEM/F12 (Lonza) supplemented with 10% FBS and OCI-LY-19 (human lymphoma cells) were cultured in RPMI 1640 (Lonza) supplemented with 10% FBS. Murine 4T1, LLC, NIH3T3 and B16F10 were cultured in DMEM supplemented with 10% FBS. RNA from human normal breast, placenta and skin were obtained from the Surgery, Obstetrics and Gynecology Department and from the Dermatology Department. Mouse RNA was purified from 8-weeks old C57Bl6 mice.