Figure 3: Substrate rigidity controls pathways of mechanotransduction involved in hepatocyte differentiation. (a) Western blot analysis of phospho-ERK1/2 and total ERK1/2 (used as a loading standard) at 3 and 24 hours after seeding on substrates with the indicated values. Images are representative of three independent experiments. (b) Flow cytofluorimetric analysis of cell cycle in RLSCs cultured at the indicated conditions for 24 hours. The values, obtained from three independent experiments, are reported as mean ± SD; , . (c) Immunofluorescence analysis of RLSCs cultured on Petri dish (CTRL), 0.4 kPa and 80 kPa for 12 hours, stained for YAP protein. The nuclei were stained with DAPI. Images are representative of three independent experiments. Scale bar: 20 μm. (d) RT-qPCR analysis of the YAP target gene, Ctgf. Data are expressed as fold change in gene expression in cells grown on 0.4 kPa and 80 kPa for 24 hours versus CTRL (arbitrary value = 1). The mean ± SD of three independent experiments is shown; .