HC11 cell differentiation and Ca2+ influx. Total mRNA was isolated and assessed for an increase in β-casein mRNA to confirm differentiation using real-time RT-PCR. Calcium assays were conducted at the same time to measure changes in basal Ca2+ influx in response to differentiation. A β-Casein mRNA levels increased in differentiated HC11 cells in comparison to proliferating cells. Data were normalized to Ppib and Actb mRNA, and are shown as relative fold expression to differentiated HC11 cells (n = 4; mean ± SD, *P < 0.05). B Basal Ca2+ influx is shown as a trace of mean fluorescence relative to baseline in proliferating and differentiated HC11 cells. C Rate of basal Ca2+ influx, calculated as the slope between 0–5 s after 2 mM Ca2+ addition, in differentiated and proliferating HC11 cells. Rate is expressed as a percentage relative to differentiated cells treated with siNT and CPA (n = 4; mean ± SEM, *P < 0.05).
Ross et al. BMC Cell Biology 2013 14:57 doi:10.1186/1471-2121-14-57