Figure 3From: A Functional Role for ADAM10 in Human Immunodeficiency Virus Type-1 ReplicationHIV-1 nuclear entry, but not completion of reverse transcription, is affected by ADAM10 down-regulation. (A) Primary human macrophages were transfected with either ADAM10 or ERBB2IP siRNA 48 h prior to infection with HIV-SF162. DNA was isolated 48 h after infection and real time PCR was used to quantitate formation of full length HIV cDNA. In order to amplify HIV cDNA with these full length primers, two template-switching events and continuous 5'LTR and gag sequences must be present on either strand, which is the last event to occur during HIV reverse transcription [79]. (B, C) Integration of HIV was significantly lower in ADAM10 down-regulated (B) primary human macrophages and (C) U373-MAGI-CCR5 cells than in control ERBB2IP down-regulated cells following infection with HIV-SF162. Genomic DNA was used to quantitate integrated HIV cDNA using real-time PCR using primers specific for integrated HIV cDNA. (D) 2-LTR circle formation in ADAM10 down-regulated macrophages was significantly less than that seen in macrophages treated with the integrase inhibitor raltegravir, and was similar to infected but untreated cells. Formation of 2-LTR circles was quantitated by real-time PCR [80]. (No Inf = No Infection, Ral = Raltegravir, Scr = Scrambled siRNA, **P < 0.01).Back to article page