Fig. 7.

The transfection efficiency and antiviral activity of TAT-P1 with DIG-3 could be increased by P1 peptide. a Transfection efficiency of TAT-P1/pLuc increased by additional ATPase inhibitor (bafilomycin A1) or P1 peptide. Before TAT-P1/pLuc (2.0 μg/0.5 μg) complex was added to 293T cells for transfection, the indicated concentrations of bafilomycin A1 (A1, nM), P1 peptide (μg per ml), or PA1 peptide (μg per ml) were added to cell culture media. Mock indicates cells treated with A1 or TAT-P1 without DNA. Data were presented as mean ± SD of three independent experiments. b Transfection efficiency of TAT-P1/pCMV-Luc increased by P1 in mouse lungs. c Representative In Vivo Imaging System image showed increased luciferase expression by P1 in mouse lungs. TAT-P1/pCMV-Luc (20 μg/5 μg) with additional P1 (20 μg, 10 μg, or 0 μg) or PA1 (20 μg) were inoculated to mouse lungs at 48 and 24 h before measuring bioluminescence signal or taking bioluminescence image. Mock indicates mouse lungs inoculated with TAT-P1 + P1 without DNA. d The RNA expression of DI-PA increased by P1 in mouse lungs. TAT-P1/DI-PA (20 μg/5 μg) with additional P1 (20 μg, 10 μg, or 0 μg) were inoculated to mouse lungs at 48 and 24 h before detecting RNA expression. Data were presented as mean ± SD of ≥3 mice. * Indicates P < 0.05 and ** Indicates P < 0.01 when normalized to Mock (a, b) or naked plasmid of DI-PA (d). e, f The prophylactic efficacy of TAT-P1/DIG-3 + P1 against A(H1N1)pdm09 virus. PBS (n = 10), TAT-P1 + P1(n = 10) or TAT-P1 + PA1 (20 μg + 20 μg, n = 5), TAT-P1/DIG-3 (20 μg/5 μg, n = 10), TAT-P1/DIG-3 + PA1 (20 μg/5 μg + 20 μg, n = 5), or TAT-P1/DIG-3 + P1 (20 μg/5 μg + 20 μg, n = 10) were intratracheally inoculated to corresponding mice at 48 and 24 h before viral inoculation. ** Indicates P < 0.01 when compared with PBS. P values were calculated by Gehan–Breslow–Wilcoxon test for survival analysis and by the two-tailed Student’s t test for analysis in a, b, d and f