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. Author manuscript; available in PMC: 2009 Dec 1.
Published in final edited form as: Nat Biotechnol. 2009 May 17;27(6):567–571. doi: 10.1038/nbt.1541

Figure 2. PTD-DRBD siRNA delivery into T cells and HUVECs.

Figure 2

(a) Flow cytometry analysis of dGFP RNAi knockdown decay kinetics of dividing Jurkat dGFP cells following treatment with GFP2 siRNA plus PTD-DRBD, Lipofection-2000 (Lipofection) or RNAiMAX (Lipofection 2), as indicated. (b) Quantitative RT-PCR analysis of endogenous GAPDH mRNA expression at 12 h post-treatment of GAPDH siRNA or GFP2 (Con) siRNA plus PTD-DRBD, GAPDH siRNA plus Lipofection-2000 (Lipofection) or RNAiMAX (Lipofection 2) in Jurkat cells, as indicated. Mean values normalized to β2 microglobulin and reported as percent of mock GAPDH control. (c) Flow cytometry histogram analysis of PTD-DRBD mediated CD4 or CD8 RNAi response at 1 day post-treatment of mouse primary T cells, as indicated. (d) Quantitative RT-PCR analysis of endogenous CD4, CD8 or CD90 mRNA expression at 12 and 24 h post-treatment of PTD-DRBD CD4 or CD8 siRNAs in primary T cells, as indicated. Mean values normalized to β2 microglobulin and reported as percent of mock control. *(P<0.05) of specific siRNA vs. control siRNA delivered by PTD-DRBD. (e) Quantitative RT-PCR analysis of endogenous GAPDH mRNA expression at 6, 12, and 24 h post-treatment of PTD-DRBD GAPDH or control siRNAs in primary HUVEC cells, as indicated. Mean values normalized to β2 microglobulin and reported as percent of mock GAPDH control. **(P<0.01) of specific siRNA vs. control siRNA delivered by PTD-DRBD. (f) PTD-DRBD cytotoxicity analysis. HUVEC cells were treated with mock (PBS), GAPDH siRNA plus PTD-DRBD or lipofection and analyzed for cytotoxicity by FACS after 24 h post-treatment with two independent means, propidium iodide and Calcein-AM. Percent indicates viable cells present in bottom, right quadrant.