Figure 7
Over-expression of P311 in C2C12 cells. (A) P311 expression, through infection with an adenoviral construct and through stable transfection with an expression plasmid, was determined over a time course. Proliferating cells were infected for 3 days in proliferation medium [PM] (3I), followed by 3 days in differentiation medium [DM] (3I 3D) or 6 days in DM (3I 6D). Stably transfected cells were incubated for 3 days in PM (PM), followed by 3 days in DM (DM 3D) or 6 days in DM (DM 6D). Results expressed as mean ± standard deviation from triplicate samples within the same experiment. (B) Adenovirus-mediated (at 0.5 MOI) expression of P311 in C2C12 cells (infected for 3 days during proliferation followed by 6 days of differentiation), viewed under fluorescence for GFP and immunostained for FLAG. P311-infected cells showed co-localisation of both proteins. (C) The viability of P311-infected and P311-transfected C2C12 cells was similar to control cells over an extended culture period, as demonstrated using a CellTiter-Blue kit (Promega). For P311 adenovirus, cells were infected in PM for 3 days (3I), followed by 3 day DM (3I 3D), or by 6 days in DM (3I 6D), or 10 days in DM (3I 10D). Stably transfected cells were grown in PM for 3 days (PM), followed by 3 day in DM (DM 3D), or 6 days in DM (DM 6D), or 10 days in DM (DM 10D). Results expressed as mean ± standard deviation from triplicate samples within the same experiment.