Effect of low virus titer-producing genes on viability of the transfected cells.

<p>HEK293T cells were transfected with gene (x-axis) expressing plasmids either alone or with viral packaging vector day after seeding. 21 h after transfection, medium was replaced with PBS containing Hoechst33342 and propidium iodide (PI), which stain DNA and dying cells respectively. Cells were scanned in blue (Hoechst, A) red (PI, B) and green (GFP, C) channels and number of objects determined in each channel. A and B represent independent object counts, data in C are expressed as proportion of GFP positive cells in the Hoechst stained population. A - Significantly reduced number of surviving cells in the well compared to untreated wells (CELLS): lipofectamine alone (MOCK) (P = 0.034); MOCK with packaging plasmids (P = 7.5E−10); all others (P≤1.1E−05) B - Significantly increased PI positive cells compared to vector: expression plasmid only BCL2L1 (P = 6.88E−05), C3ORF1(P = 0.03), MTCH1(P = 4.66E−05), PNMAL2 (P = 0.01) P values determined using Aspin-Welch test; Bars = mean of 4 wells, error bars = SD, CELLS-untreated well, MOCK-wells where expression plasmid has been omitted. AWAT2, and vector were high virus titer producing in previous experiments.