Fig. 7.
(A) Jurkat cells were stained with anti-CD71 antibody (TfR) or isotype antibody. The median fluorescence intensity (MFI) of antibody-stained of cells was quantified by flow cytometry. (Data points indicate mean +/− SD, n=2; One-way ANOVA, ***, p<0.005; Yellow bar: MFI of Jurkat cells stained with CD71 antibody, green bar: MFI of Jurkat cells stained with isotype antibody, orange bar: background MFI of Jurkat not stained with antibody ). (B) Cellular uptake in Jurkat cells of Tf-PEI, PEI and lipofectamine 2000 (LF) polyplexes/lipoplexes prepared with 50 pmol AF488 siRNA at different N/P ratios (5, 10, 20) was quantified by flow cytometry (Data points indicate mean +/− SD, n=2; One-way ANOVA, ***, p<0.005; Orange bars: control groups include a “blank” group of untreated cells, the “siRNA” group are cells only treated with free siRNA, the “LF” group represents cells treated LF lipolexes, yellow bars: cells treated with Tf-PEI polyplexes, green bars: cells treated with PEI polyplexes). (C) GAPDH gene knockdown efficiency of Tf-PEI (N/P 15) was compared with PEI polyplexes (N/P 15) and LF lipoplexes. GAPDH expression was normalized with beta-actin expression and quantified by real time-PCR. (Data points indicate mean +/− SD, n =3; One-way ANOVA, **, p < 0.01, ***, p<0.005; Orange bar: the blank group represents GAPDH expression of untreated cells, yellow bars: GAPDH expression of cells treated with polyplexes/ lipoplexes containing siRNA against GAPDH (siGAPDH), green bars: negative control group, GAPDH expression of cells treated with polyplexes/ lipoplexes containing scrambled siRNA (siNC) ).