Modulation of Gene Expression by Substrate Stiffness via Regulation of Histone H2B Mono-ubiquitination

Mechanotransduction plays a critical role in regulating cell growth, proliferation, and metabolism. Cells respond to mechanical signals, which ultimately induces gene expression. However, the underlying mechanisms of this gene regulation remain largely unclear. In previous research, we discovered that the ubiquitin-conjugating enzymes UBE2A and UBE2B translocate between the cytoplasm and nucleus in a force-dependent manner. Additionally, we confirmed that UBE2A and UBE2B ubiquitinate histone H2B at lysine 120 (K120) when cells are cultured on a stiff substrate. In this study, we employed Chromatin Immunoprecipitation (ChIP) to investigate the distribution of H2B mono-ubiquitination in human skeletal muscle (hsSKM) cells cultured on substrates of varying stiffness—soft (0.2 kPa) and stiff (64 kPa). Overall design: chromatin immunoprecipitation for H2BK120ub of hsSKM cell cultured on 64kPa or 0.2kPa hydrogel