Fig 1: Evaluation of PTEN effects on BTG2 expression and on cell growth in vitro and in vivo in human bladder cancer cells. (A) The expressions of PTEN in three bladder cancer cell lines were determined by immunoblotting assays. The data are expressed as the intensity of the PTEN protein bands/β‐actin (± SE; n = 3) relative to the RT4 cells. The mRNA expressions of PTEN and BTG2 in PTEN‐knockdowned RT4 (RT4_shPTEN; white bars) cells, mock‐knockdowned RT4 (RT4_shCtrl; black bars) cells (B), PTEN‐overexpressed T24 (T24‐PTEN; white bars) and mock‐overexpressed T24 (T24‐DNA; black bars) cells (C) were determined by RT‐qPCR assays. Data are expressed as mean‐fold (± SE; n = 3) of the target genes relative to mock‐treated group. (D) BTG2 report vector was co‐transfected with various concentrations of PTEN expression vector into T24 cells for 72 h. Data are expressed as the mean percentage ± S.E. (n = 6) of luciferase activity relative to mock‐transfected groups. (E) The rates of cellular proliferation in T24‐DNA cells and T24‐PTEN cells were analyzed by 3H‐thymidine incorporation assays. (F) The rates of cellular proliferation in RT_shCtrl cells and RT4_shPTEN cells were analyzed by 3H‐thymidine incorporation assays. (*P < 0.05, **P < 0.01).
Fig 2: Effects of PTEN modulation on downstream signal transductions and BTG2 in human bladder cancer cells. (A) The expressions of PTEN, pAKTs473, pAKTt308, AKT, pGSK3b, GSK3b, pmTOR, mTOR, P70S6K, pP70S6K, and BTG2 in T24‐DNA and T24‐PTEN cells (left), and in RT4_shCtrl and RT4_shPTEN (right) were determined by immunoblotting assays. (B) RT4 cells were treated with various dosages of VO‐OHpic trihydrate. Expressions of PTEN, Akt, p‐Akt (t308 and s473), BTG2, and β‐actin were determined by immunoblotting assays. Expressions of BTG2 mRNA in RT4 (C) and PTEN‐overexpressed T24 (D) cells following various concentrations of VO‐OHpic trihydrate treatments were determined by RT‐qPCR assays. (E) The BTG2 reporter vector‐transfected T24‐PTEN cells were treated with various concentrations of VO‐OHpic trihydrate for 24 h. Data are expressed as the mean percentage ± S.E. (n = 6) of luciferase activity relative to solvent‐control groups. (**P < 0.01).
Supplier Page from Sino Biological, Inc. for Human PTEN Gene ORF cDNA clone expression plasmid