Expression of mA3 transcripts and protein in the spleens of B6 and BALB/c mice.

<p>(A) Schematic representation of the mouse APOBEC3 exon composition. The exons are indicated by numbers and their relative sizes are depicted. The arrows indicate the locations of the PCR primers used in the following experiments. (B and C) RT-RCR and quantitative real-time PCR (qPCR) analyses showing splicing patterns and quantities of mA3 mRNA in B6 and BALB/c spleen cells. The primers utilized were c-d for assays shown in (B) and a–b for those in (C). 5+, exon 5-containing mA3; Δ5, mA3 lacking exon 5. Actin was used as an internal control. Quantitative real-time PCR data show averages of three reaction wells and SD. (D) Immunoblot detection of mA3 protein expressed in mouse spleen cells. The spleen cell extracts were tested for their total protein content by Bradford assays and the same amount of protein was loaded in each lane. Anti-mouse APOBEC3 NT antibody reactive with an N-terminal epitope was first pre-absorbed with the spleen lysate prepared from an mA3-knockout (KO) mouse, and was used as the primary antibody. Mouse actin was used as a loading control. Three independent experiments were done for all assays, and the results were consistent with the representative ones shown here.