Figure 5.

Increased basal expression of α-SMA in pten^−/− cells is due to increased gene expression and transcription. (A) Two separate aliquots of wt and pten^−/− cells were cultured in serum-free media in the presence or absence of TGF-β (2 ng/ml) for 24 h. Whole-cell lysates were evaluated by Western blot for α-SMA expression. To verify equal protein loading, the membrane was stripped and reprobed for β-tubulin. (B) The wt and pten^−/− cells were cultured in serum-free media for 24 h and RNA was isolated. α-SMA gene expression was evaluated by semiquantitative real-time PCR using glyceraldehyde phosphate dehydrogenase as an internal control. Results are pooled data from two separate experiments performed in triplicate.