REAF is downregulated in response to HIV-1 infection and restriction requires reverse transcription. (A) Time course of REAF mRNA following challenge with HIV-189.6. qPCR results from analysis of cDNA are normalised to β-actin mRNA levels. (B) REAF protein levels are downregulated following challenge with HIV-189.6. Treatment with MG132 (10 μM) prevents the degradation of REAF protein. Tubulin is included as a loading control. Only the lower band of REAF (80 kDa) is shown – not all bands are detected in all experiments. (C) Following treatment with AZT (100 nM), challenge with HIV-189.6 no longer causes depletion of REAF at 1 hr pi. GAPDH is included as a loading control.