Journal: Stem cell research
Article Title: Vpx mediated degradation of SAMHD1 has only a very limited effect on lentiviral transduction rate in ex vivo cultured HSPCs
Figure Lengend Snippet: Vpx+ virus, dN and dNTP treatments result in minor increases in the dNTP pool of HSPCs. MDMs (A, C and E) and freshly isolated HSPCs (B, D and F) were pretreated for 2 h with Vpx+/− viruses, dNs or dNTPs, and cultured for 3 days before cell lysis. Quantification of dNTPs was done by a single nucleotide incorporation assay (see “Material and methods” section). Fold induction was calculated based on the absolute values of vero- vs mock-treated samples (C–F). White dots represent Vpx− virus-pretreated samples, black dots represent Vpx+ virus-pretreated samples (A–B). White bars represent dN-pretreated samples, black bars represent dNTP-pretreated samples (C–F). Data are from three donors and bars represent mean ± SEM. Statistical analysis was done using two-tailed paired t-test.
Article Snippet: THP-1 cells transduced with lentiviral vectors encoding SAMHD1 shRNA were selected by culturing in a medium containing 0.8 μg/ml puromycin. dNs consisted of a mixture of dA (D8668), dC (D0776), dG (D0901) and dT (T1895; all from Sigma-Aldrich). dNTPs (R0192) were purchased from Fermentas (Glen Burnie, MD).
Techniques: Isolation, Cell Culture, Lysis, Two Tailed Test