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. 2016 May 27;6:26616. doi: 10.1038/srep26616

Figure 1. Silencing of SAMHD1 increases HBV replication in resting HepG2.2.15 cells.

Figure 1

(a) SAMHD1 or actin protein levels were detected in cell lysates from SAMHD1 knockdown or control HepG2.2.15 cells by Western blotting. (b) Phosphorylation on residue T592 of SAMHD1 and cyclin B1 expression were detected by Western blotting of lysates from control shRNA HepG2.2.15 cells cultured for 3 days in complete versus serum-free medium. (c-e) HepG2.2.15 cells stably expressing two different SAMHD1 shRNAs or a control shRNA were cultured in serum-free medium for 3 days. (c) Luciferase activity (relative luciferase units, RLU) was detected in cells 24 hours post infection with a single-round HIV-1-based lentiviral vector that encoded luciferase. (d) Equal cell growth and viability between different shRNA cell lines was assessed using ATPLite. (e) The amount of HBV DNA in the supernatant was determined by qPCR and normalized to the control shRNA. In (c,d), the mean ± standard error mean (SEM) of three biological replicates of one representative experiment is depicted. In (e), the fold-changes to negative control were calculated for each individual experiment based on the median of three biological replicates, each measured in three technical replicates. The mean ± SEM of the fold-changes of four independent experiments is depicted. Statistical significance was determined using a one-way ANOVA with multiple comparisons according to Dunnett (*p < 0.05; ****p < 0.00005).