Figure 2.

Up-regulation of CDK inhibitor proteins by v-Src-induced tyrosine phosphorylation. (a) HeLa S3/TR/v-Src-wt cells were cultured with or without 1 µg/ml Dox for 24 h, and whole cell lysates were analyzed by Western blotting (WB) using anti-p21, anti-p16, anti-Src (N-16), and anti-actin antibodies. (b–d) HeLa S3/TR/v-Src-wt cells were synchronized as depicted in Fig. 1a. (b) Whole cell lysates were analyzed by Western blotting using anti-p21, anti-p16, anti-Src (N-16), and anti-actin antibodies. Asy stands for asynchronous HeLa S3/TR/v-Src-wt cells. (c) Synchronized cells at the G₁/S boundary were released for the indicated times with 1 µg/ml Dox, and whole cell lysates were analyzed by Western blotting using anti-p21, anti-Src (GD11), and anti-actin antibodies. (d) Synchronized cells at the G₁/S boundary were cultured for 6 h with 1 μg/ml Dox in the presence or absence of 10 μM SU6656. Whole cell lysates were analyzed by Western blotting using anti-p21, anti-Src (N-16), and anti-actin antibodies. (e) HeLa S3/TR/v-Src(K295M) cells were cultured with or without 1 µg/ml Dox for 24 h. Whole cell lysates were analyzed by Western blotting using anti-p21, anti-Src (N-16), and anti-actin antibodies.