Figure 6.

Cell Viability Screen of Exon Replacement Constructs in HIV-Producing Cells

293T cells were transfected with pNL4.3 and sequentially transfected with RNA trans-splicing constructs the following day. Cells were treated with one or two doses of GCV at a concentration of 100 μM, and cell viability was measured by the MTT assay 6 days after the initial transfection. Ctrl refers to untransfected 293T cells, pVAX-1 (empty vector backbone) was used as a negative control, and HSV-tk (full length HSV-tk) was included as a positive control. (A) 3′ exon replacement in HIV-producing 293T cells in the presence or absence of GCV. (B) Cell viability screen of 5′ exon replacement constructs in HIV-producing 293T cells in the presence or absence of GCV. BD-A3 (top left), BD-A5 (top right), BD-A7 (bottom left), and BD-A8 (bottom right).The data in (A) and (B) represent the average mean of representative experiments performed in multiple wells (three to eight wells). The error bars represents the SD of the average percentage cell viability. p values (two-tailed) are shown for “t test: two sample assuming unequal variances”; *p ≤ 0.05), where statistical significance was observed in each of three independent experiments.