Figure 8

Compromising Paks suppresses HTLV-1 proviral gene transcription. (A) RNAi depletion of endogenous Pak1 and Pak3 decreases the expression of HTLV-1 proviral mRNA transcripts. HeLa cells were transfected with siPak1 or siPak3 to knockdown endogenous Pak1 and Pak3, respectively. After 24 h, cells were transfected with HTLV-1 molecular clone pX1MT. Cells were harvested 48 h after pX1MT transfection. Total RNA was extracted by Trizol and cDNA was synthesized. Quantitative RT-PCR was performed to analyze the relative levels of proviral Tax, Gag, Env, Pak1, Pak4 and β-globin transcripts. siGFP was used as a negative control. Relative expression levels were analyzed by normalizing to β-globin and fold difference represent 2^–ΔΔCT. (B) RNAi depletion of Pak1 and Pak3 decreases the expression of Tax transcript in HTLV-1-transformed T cells. MT4 cells were transfected with siGFP, siPak1 and siPak3. siGFP was used as a negative control. After 72 h of siRNA transfection, total RNA was extracted and quantitative RT-PCR was performed. (C) Proliferation of MT4 cells was unaffected by siPak1/3 as measured by the MTT assay.