FIG. 4.
Expression of HIV-1 Nef in CD4C/HIV-Nefallele Tg mice. Thymuses from different founder lines with each of the CD4C/HIV-Nefallele transgenes were analyzed. (A) Northern blot analysis of HIV-1 RNA. Total RNAs from thymuses were hybridized with a 32P-labeled HIV-1-specific probe. The blots were stripped and rehybridized with a 32P-labeled actin DNA probe. Also shown below are the semiquantitative values for the levels of expression of each Nef allele relative to NefNL4-3(WT) (value of 1), measured in Northern blot or quantitative RT-PCR (Q-RT-PCR) analysis, as described in Materials and Methods. (B) Western blot analysis of protein extracts (100 μg) from thymuses of 1-month-old Tg and non-Tg littermates, using the following anti-Nef antibodies: rabbit polyclonal antibody produced in our laboratory (Lab) or monoclonal EH1 or 6.2 antibody. NefNL4-3(WT) represents the gene carried by the CD4C/HIVMutG (F27367) Tg mice previously reported (34) and used as a positive control. The membranes were stripped and reanalyzed with anti-actin antibody.