FASN regulates bioavailability of VEGF-A via alteration of activity of MMPs. (A) Immunoblot for MMP-9 in CRC cell lines with stable knockdown of FASN (top arrow-proenzyme, bottom arrow-activated enzyme). (B) Zymography and densitometry analysis for MMP9 in CRC cell lines with stable knockdown of FASN. (C) Expression of MMP-9 in CRC cell line assessed by confocal microscopy, ×60 magnification. (D) IF staining of HCT116 and HT29, NTC and FASNsh, orthotopic tumors for VEGF-A (red), MMP-9 (green) and 4′,6-diamidino-2-phenylindole (DAPI) (blue). Arrows point at VEGF-A associated with ECM. (E) Expression of VEGF-A in HT29 cells treated with MMPs inhibitors for 24h.