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. Author manuscript; available in PMC: 2005 Sep 13.

Published in final edited form as: J Biol Chem. 2005 Apr 25;280(27):25788–25801. doi: 10.1074/jbc.M413594200

Fig. 10 — Transient expression of a p21 construct, phosphomimetic at the Mirk phosphorylation site, increases the viability of C2C12 cells. C2C12 myoblasts were transfected with Flag-p21 wild-type, Flag-p21-S153A, Flag-p21-S153D, or vector for 4 hours, allowed to express for 24 hours (0 point) then swtiched to serum-containing growth medium for 0–13 days. A. Each construct was expressed for only about 1 day in growth medium following the transfection and initial 24 hour expression period, as determined by western blotting for the Flag epitope. The zero time points for each construct were examined in parallel to the zero time points for the other two constructs on each blot as internal controls. Ct, cross-reacting protein used as a loading control. B. After 24 hours, the cells were trypsinized and plated at either 500 or 1000 cells in 100 mm tissue culture dishes, each in triplicate. Data shown is the mean of two such experiments, normalized to the number of colonies per 1000 cells plated.

Fig. 10 — Transient expression of a p21 construct, phosphomimetic at the Mirk phosphorylation site, increases the viability of C2C12 cells. C2C12 myoblasts were transfected with Flag-p21 wild-type, Flag-p21-S153A, Flag-p21-S153D, or vector for 4 hours, allowed to express for 24 hours (0 point) then swtiched to serum-containing growth medium for 0–13 days. A. Each construct was expressed for only about 1 day in growth medium following the transfection and initial 24 hour expression period, as determined by western blotting for the Flag epitope. The zero time points for each construct were examined in parallel to the zero time points for the other two constructs on each blot as internal controls. Ct, cross-reacting protein used as a loading control. B. After 24 hours, the cells were trypsinized and plated at either 500 or 1000 cells in 100 mm tissue culture dishes, each in triplicate. Data shown is the mean of two such experiments, normalized to the number of colonies per 1000 cells plated.