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Figure 3 | Retrovirology

Figure 3

From: ADAR1 enhances HTLV-1 and HTLV-2 replication through inhibition of PKR activity

Figure 3

ADAR1 expression enhances HTLV-1 and HTLV-2 protein expression. (A, B): Jurkat or (C, D) PBLs were transfected with (A, C) 2 μg of pACH or (B, D) pH6neo molecular clones together with 1 μg of a plasmid encoding ADAR1 and 2 μg of (A, C) HTLV-1-LTR or (B, D) HTLV-2-LTR-luc plasmids. Forty-eight hours later, luciferase activity was measured and results normalized by protein concentration as determined by the Bradford method and calculated as fold change compared to cells that were not transfected with ADAR1 plasmid arbitrarily set to 100%. (A, B): Data are the mean ± standard deviation (SD) from 3 independent experiments. Asterisks indicate statistically significant differences between treated and untreated cells (paired Student t test, *p < 0.05). (C, D): Data are representative of two different experiments obtained with two different blood donors. (E, F): 293T-LTR-GFP cells were transfected with 1 μg of pACH (HTLV-1) or pH6neo (HTLV-2) molecular clones together with 125 ng of an ADAR1 expression plasmid or of the backbone vector. Forty-eight hours later, cells were visualized and pictures taken with a Zeiss Axio Vert.A1 imaging microscope. (G, H, I, J): 293-T cells were transfected with 4 μg of (G, I) pACH (HTLV-1) or (H, J) pH6neo (HTLV-2) molecular clones and increasing amount (0, 5, 50, 500 ng) of an ADAR1 expression plasmid. (G, H): Western blot analyses were performed on 60 μg of proteins obtained from whole cell extracts using anti-ADAR1, anti-p24gag, anti-Tax1, anti-Tax2 and anti-actin antibodies. (I, J): p19gag was quantified in the culture supernatants from cells transfected as in (G, H). Data are represented as fold change compared to the p19gag values obtained for cells that were not transfected with the ADAR1 plasmid arbitrarily set to 100%.

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