FIG. 7.

Antisense oligonucleotides against p21 inhibit the induction of p21 and Akt during myogenic differentiation. (A) C2C12 cells were cultured in standard differentiation medium (−) or in differentiation medium containing Lipofectin with 0.5 μM p21-antisense (p21-AS) or control (Cont) oligonucleotides for 24 h. After 24 h of incubation, cells were harvested and cell lysates (20 μg protein) were immunoblotted with anti-Akt, anti-p21, and anti-Cdk4 antibodies. Representative data from one of five assays are shown. (B) Quantitative analysis of reduced Akt expression. The intensities of the Akt and Cdk4 bands in immunoblots from five independent experiments were quantified by gel densitometry. The ratios of Akt band intensity relative to Cdk4 band intensity were calculated and normalized to the ratios of band intensities in parallel cultures that were incubated in differentiation medium with no oligonucleotide or Lipofectin. Data are shown as mean ± SEM.