QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 2. Downregulation of endogenous Twist-1 using RNAi methods enhances BMP-induced transcriptional activity mediated by Smads. (A) C3H10T1/2 cells were transfected with either Twist-1-specific siRNA (siTwist-604, siTwist-691, siTwist-645, siTwist-481) or control siRNA (Scrambled) as described in the Materials and Methods. Total RNA was extracted 24 hours later, and northern blot analysis was performed using a Twist-1 cDNA probe. (B) C3H10T1/2 cells were transfected with either siTwist-691 or control siRNA (Scrambled). Cells were transfected with 3GC2-Lux and TK-Renilla luciferase 24 hours after siRNA transfection. At 12 hours after second transfection, cells were treated, with BMP (300 ng/ml) or left untreated, for 12 hours. Cells were lysed and luciferase activity was assayed (^*P<0.01). The mean value of firefly luciferase and Renilla luciferase activity in the scrambled sample without BMP was approximately 2,000 and 34,000 RLU (relative light unit), respectively. (C) C3H10T1/2 cells were transiently transfected with either 90 pmol of siTwist-691 or Scrambled in combination with 100 ng of 3GC2-Lux luciferase construct, Smad1, Smad4 and BMPR-IB(QD). Cells were lysed and luciferase activity was assayed 24 hours after transfection (^*P<0.01). The mean value of firefly luciferase and Renilla luciferase activity in the scrambled sample without BMPR-1B(QD) was approximately 411,000 and 30,000 RLU, respectively.

Return to article