Polyacrylamide hydrogels (PAGs) were cast onto No. 0 glass bottom 35 mm culture dishes (MatTek P35G-0-20-C) using a previously described method and formulation [13 (link), 51 (link)]. Cast gels were then coated using sulfo-SANPAH (Thermo Fischer Scientific, Waltham, MA) as previously described (Bangasser et al. [13 (link)], Wang and Pelham [51 (link)]). In this study, Type I Collagen (354236, Corning, Corning, NY) or anti-CD44 antibody α-CD44 MAB (BDB553131, BD Biosciences, San Jose, CA) were used to coat PAGs. In general, 200 μg/mL Col I solution was used and 1-300 μg/mL α-CD44 MAB solution was used depending on the experiment. The Young’s Modulus of the PAGs was determined using the methods described by Bangasser et al.. 0.7, 4.6, 9.3, 19.8, 98.5, 195 kPa stiffnesses were used in this study, which were obtained by varying the cross-linker and polymer concentrations as previously described [13 (link)].
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