Figure 3.

Verification of knockdown of STMN1 gene expression in esophageal cancer cell line by lentivirus-mediated RNA interference. (A) Western blot analysis of STMN1 protein in lysates of either untreated (control) or transfected with a Non-silencing shRNA (scrambled sequence) and transfected with a specific STMN1 shRNA. ACTIN expression was used as a loading control. (B) Densitometry analysis of A normalized to Actin, Laser densitometric analysis of protein bands was performed, and the ratio of stathmin expression to Beta-Actin expression (STMN:Beta-Actin) was determined and normalized. (C) Total RNA isolated from distal esophageal adenocarcinoma cells before and after silencing stathmin gene expression using STMN1 shRNA were analyzed with real-time RT–PCR. There was no significant difference between controls and non-silencing shRNA (scrambled sequences). Data are expressed as percentage change (Means ± S.D.) compared with controls and represent four independent experiments. (P < 0.05 vs Non-silencing shRNA, one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparion).