800cw or 680cw ir dye

Whole cell lysates or concentrated conditioned media were separated on SDS-PAGE gels (7%, 10%, or 4-15% gradient gels, Criterion Gel System; Bio-Rad, Hercules, CA) via standard techniques. Proteins were transferred to nitrocellulose membranes using a Bio-Rad Trans-Blot Turbo system and blocked in Odyssey Blocking buffer (Li-Cor Biosciences, Lincoln, NE) prior to overnight incubation in a 1μg/mL solution of primary antibody of interest. Primary antibodies included: caveolin-1, collagen I, collagen III (Abcam, Cambridge, MA), Cavin-1, Cavin-3, fibronectin, matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP9), tissue inhibitor of metalloproteinase 1 (TIMP1), and tissue inhibitor of metalloproteinase 2 (TIMP2) (Santa Cruz Biotechnology, Santa Cruz, CA). Secondary antibodies were conjugated to 800CW or 680CW IR dye (Li-Cor Biosciences) and imaging was performed using a Li-Cor OdysseyXL system prior to quantification via densitometry. GAPDH expression was used to normalize cell lysate protein concentrations. Concentrated conditioned media was normalized to cell lysate protein concentrations as determined by Lowry protein assays.