Skip to main content
Figure 3 | Retrovirology

Figure 3

From: Identification and characterization of naturally occurring splice variants of SAMHD1

Figure 3

Interaction of Vpx and SAMHD1 splice variants. (A) HeLa cells were transfected with pcDNA-SAMHD1 WT (1 μg), Δ14 (5 μg), or Δ8-9 (4 μg) as indicated, either without Vpx (lanes 1, 4, 7) or with increasing amounts (0, 0.25, 0.5 μg) of pCMV-Vpxmac239. Whole cell extracts were prepared 24 h after transfection and separated on 10% (SAMHD1, actin) or 15% (Vpx) SDS-PAGE. Immunoblot analysis was performed with rabbit polyclonal antibodies specific to SAMHD1 and SIVmac239 Vpx. The SAMHD1 blot was subsequently reprobed with a rabbit polyclonal antibody to actin. (B) Vpx interacts with SAMHD1 splice variants. HeLa cells were transfected with pcDNA-HA-SAMHD1 WT (2 μg), Δ14 (5 μg), or Δ8-9 (3 μg) together with 0.5 μg of pCMV-Vpxmac239. Total amounts of transfected DNA were adjusted to 5.5 μg with empty vector DNA as appropriate. MG132 (10 μM final concentration) was added to the samples 20 h post-transfection and cells were lysed 4 h later and processed for immunoprecipitation with HA-beads as described in Methods. Immunoprecipitated proteins (IP: α-HA) together with input lysate (input) were separated by SDS-PAGE and immunoblot analysis was performed with antibodies to SAMHD1 or Vpx as indicated.

Back to article page