(a-c) Brightfield images of PCs cultured on RGD-coated glass (E>2GPa), 10kDa (E: 83.8kPa), or 20kDa (E: 54.7kPa) hydrogels. Scale bars are 80 microns. (d) Quantification of cell size across all three substrates. *p<0.05, ***p<0.001 as determined by one-way ANOVA. (e-h) Phalloidin (red) and DAPI (blue) staining on sub-confluent PCs on glass (e), 10kDa (f), and 20kDa (g) gels and quantification of the actin intensity (h). ****p<0.0001 as determined by unpaired t-test. (i-l) NG2 (red) and DAPI (blue) staining on PCs on glass (i), 10kDa (j), and 20kDa (k) gels with representative flow cytometry dot plots shown below each image. Quantification of intensity by flow cytometry (l). (m-o) PDGFRβ (green) and DAPI (blue) staining on PCs on glass (m), 10kDa (n), and 20kDa (o) gels with representative flow cytometry dot plots shown below each image. Quantification of the intensity of expression (p). (q-s) Connexin43 (green) and DAPI (blue) staining on PCs on glass (q), 10kDa (r), and 20kDa (s) gels with representative flow cytometry dot plots shown below each image. Quantification of staining intensity by flow cytometry (t). Scale bars in fluorescent images are 10 microns.