Figure 1

Identification and characterization of SAMHD1 splice variants. (A) Schematic representation of the SAMHD1 protein and the SAMHD1 splice variants cloned from PMA-treated THP-1 cells. (B) Location of the primers used for the specific amplification of SAMHD1 splice variants. Δ14 primers consist of a forward primer (F14) located in exon 10 and a reverse primer (R14) spanning the junction of exons 13 and 15. The resulting PCR product is predicted to be 462 bp long. Δ8-9 primers consist of a forward primer (F8/9) spanning the junction of exons 7 and 10 and a reverse primer R8/9 in exon 15, resulting in a predicted PCR product of 711 bp. (C) cDNA pools were generated from THP-1 cells treated with 100 nM PMA for 24 h and used as template for a PCR reaction using Taq polymerase and the primer sets described in panel B. Plasmid controls consisted of 1 ng of pcDNA-SAMHD1 WT, Δ14, or Δ8-9 used as template in the PCR reaction, as indicated. (D) HeLa cells were transfected with 5 μg each of pcDNA-SAMHD1 WT, Δ14, or Δ8-9 as indicated. Total cell extracts were prepared 24 h later and separated on 10% SDS-PAGE. Immunoblot analysis was performed with rabbit polyclonal antibodies specific to SAMHD1 (SAM416) and actin, as indicated.