Figure 1

Analysis of single VLP treatment in MDMs. (A) MDMs were treated with Vpx- or Vpx + VLPs starting at day 0 and cell lysates were collected every 24 h until day 14. Immunoblots were probed for SAMHD1 or GAPDH, loading control. One representative MDM donor of three is displayed. (B) Quantitation of SAMHD1 expression (20 μg) by Western blot analysis is shown for the three independent MDM donors. Data are graphed by setting SAMHD1 expression level at day 0 to 100%. Two-way ANOVA was done with Bonferroni post-test analysis to determine significance, *** = P < 0.001; ** = P < 0.01. (C) Western blot analysis using 60 μg total protein, 3× loading as compared to (B). SAMHD1 level of expression is below 3% even with greater loading. (D-G) HIV RT-based dNTP assay was used to determine the levels of dATP (D), dGTP (E), dCTP (F) and dTTP (G). Data are graphed for the 15 days that the samples were collected. Two-way ANOVA analysis was performed and significant differences denoted on the graphs for dATP and dGTP.