Figure 2
From: HIV-1-encoded antisense RNA suppresses viral replication for a prolonged period
Antisense-specific RT-PCR of HIV-1 RNA. (A) A diagram describing the strand-specific RT-PCR method for HIV-1 asRNA. RNA samples were reverse-transcribed by an antisense-specific primer with a 5′ tag sequence (Tag-RT-primer). The synthesized cDNA samples were then amplified by PCR with a primer corresponding to the tag sequence (Tag primer) and a primer complementary to the target cDNA (PCR primer). (B) Detection of asRNAs in MAGIC-5A cells infected with HIV-1NL4-3. The top panel describes the position of R1–R5 region. Lower panels show the results of agarose gel electrophoresis of products of the antisense-specific RT-PCR targeting R1–R5 regions. PCR products were detected corresponding to all 5 regions studied. Lane 1, MAGIC-5A without virus; Lane 2, MAGIC-5A with HIV-1NL4-3; Lane 3, no RTase control; Lane 4, no RT primer control (a control for endogenous priming); Lane 5, PCR products with conventional primer pairs with cDNA samples synthesized by random primers (for testing sense and antisense RNA expressions); Lane 6, positive control (amplified from pNL4-3 plasmid DNA).