Bb 94

Dulbecco's modified Eagle medium (DMEM), MEM non-essential amino acids and 0.25% trypsin/EDTA solution were purchased from Invitrogen (Gaithersburg, MD). Platelet-derived growth factor BB isotype (PDGF), GM6001 (also known as galardin or ilomastat) were obtained from Millipore (Billerica, MA). BB-94 (also known as batimastat) was purchased from Tocris (Bristol, UK). Fetal bovine serum (FBS), fatty acid-free and fraction V bovine serum albumin (BSA), RPMI vitamin mix, HEPES, DMSO, thymidine and Sodium bicarbonate were obtained from Sigma-Aldrich (St. Louis, MO). Penicillin, streptomycin, and amphotericin B were obtained from Lonza inc. (Walkersville, MD). Type I rat tail collagen was purchased from BD Biosciences (Bedford, MA). PureCol® Type I bovine collagen was purchased from Advanced BioMatrix, Inc. (San Diego, CA). Alexa Fluor 488 and Propidium Iodine (PI) were obtained from Molecular Probes, Inc. (Eugene, OR). Collagenase D from Clostridium histolyticum and RNase (DNase free) were purchased from Roche (Indianapolis, IN). Rabbit eyes were purchased from Pel Freez (Rogers, AR). Human glu-plasminogen was purchased from Haematologic Tech (Essex Junction, VT). Glass bottom dishes were purchased from MatTek (Ashland, MA). MT1-MMP mouse-anti-human monoclonal antibody (C9, Santa Cruz, TX)

Gelatin-coated coverslips were prepared as described using Oregon green 488-labeled gelatin (Life Technologies) at a 1:8 ratio with unlabeled gelatin (11 (link)). Cells were plated on gelatin-coated coverslips and incubated for 8 hours. Alternatively, cells were incubated overnight in the presence of 2 μM BB-94 (Tocris). The following day, BB-94 was washed out and the cells were incubated for 8 hours before fixation and staining.