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. Author manuscript; available in PMC: 2011 Jul 30.

Published in final edited form as: Biochem Biophys Res Commun. 2010 Jun 25;398(3):395–399. doi: 10.1016/j.bbrc.2010.06.080

Fig. 1 — Knockdown of p70S6K1 inhibited VEGF protein expression and VEGF transcriptional activity. (A) OVCAR-3 cells stably expressing p70S6K1 siRNA (sip70S6K1) or scrambled control (scrambled ctrl) were cultured to 100% confluence. The cells were changed to fresh complete medium. Aliquots of cell supernatant were analyzed for VEGF protein levels by ELISA. (B) Cells were seeded in 12 well plates and cultured to 40–50% confluence, then transfected with VEGF promoter reporter pGL-Stu1, pCMV-β-galactosidase plasmid (β-gal), sip70S6K1 and wild-type HIF-1α plasmids at the concentrations as indicated. The empty vector was added to make total transfected DNA to 1.15 µg. Relative Luc activity was calculated as the ratio of Luc/β-gal activity. (C) The similar experiment was performed using pMAP11wt reporter as described for pGL-Stu1 reporter. (D) The pMAP11 mutant reporter was used to perform the similar experiment. *, indicates the significant difference (p<0.05).

Fig. 1 — Knockdown of p70S6K1 inhibited VEGF protein expression and VEGF transcriptional activity. (A) OVCAR-3 cells stably expressing p70S6K1 siRNA (sip70S6K1) or scrambled control (scrambled ctrl) were cultured to 100% confluence. The cells were changed to fresh complete medium. Aliquots of cell supernatant were analyzed for VEGF protein levels by ELISA. (B) Cells were seeded in 12 well plates and cultured to 40–50% confluence, then transfected with VEGF promoter reporter pGL-Stu1, pCMV-β-galactosidase plasmid (β-gal), sip70S6K1 and wild-type HIF-1α plasmids at the concentrations as indicated. The empty vector was added to make total transfected DNA to 1.15 µg. Relative Luc activity was calculated as the ratio of Luc/β-gal activity. (C) The similar experiment was performed using pMAP11wt reporter as described for pGL-Stu1 reporter. (D) The pMAP11 mutant reporter was used to perform the similar experiment. *, indicates the significant difference (p<0.05).