Figure 5

siRNA reveals the contribution of Sp1, Sp3 and HDACs1, 2, 3 to NRP-1 expression. Sp1 and Sp3 siRNA were screened for functionality and a single siRNA to each transcription factor was selected for further study. (A) Efficacy and specificity of Sp1 and Sp3 knock-down were shown by immunoblotting protein extracts from transfections and controls 48 hr after transfection. (B) The levels of NRP-1 transcript relative to negative siRNA transfected cells were determined by qRT-PCR using the comparative ΔΔCt method of quantification ns = no significant difference * = P < 0.05, ** = P < 0.01 Student's t-test compared to negative siRNA sample. n = 3 replicates. Error bars represent mean ± s.e