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Figure 3 | Retrovirology

Figure 3

From: hnRNP E1 and E2 have distinct roles in modulating HIV-1 gene expression

Figure 3

hnRNP E1 and E2 overexpression have only limited effects on HIV-1 RNA levels. 293 cells were transfected with eithermycE1 (E1), mycE2 (E2) or CMVmyc (myc) along with provirus HxBruR-/RI- and CMV PLAP. 48 hrs post transfection, cells were lifted in 1×PBS and total RNA isolated. Northern and RT-PCRs were performed. (A) Northern analysis of the effect of mycE1/E2 overexpression on levels of 2 kb, 4 kb and 9 kb classes of HIV-1 RNA. Probe derived from the HIV-1 LTR was used to detect all 3 classes of viral RNA; viral RNAs (9, 4, 2 kb) are indicated. Blots were stripped and reprobed for GAPDH RNA to confirm equal loading. At right, a summary of 3 sets of experimental results quantitated using Image Quant v5.2. Figures are normalized to GAPDH with the results for the myc control vector set at 1 for each RNA class. Error bars show standard deviation. An asterisk denotes 95% confidence level that the difference between the sample and the myc control is significant. (B) RT-PCR showing effects of mycE1 and mycE2 on the 2 kb species of HIV-1 RNA. Reporter plasmids and identity of amplicons are indicated. At right, a summary of 3 sets of experimental results quantitated using Image Quant v5.2. Shown is the percentage of each species of the total RNA for each lane. For each species of RNA the myc value was set at 1. Error bars show standard deviation. An asterisk denotes 95% confidencelevel that the difference between the sample and the myc control is significant. (C) RT-PCR showing effects of mycE1 and mycE2 on the 4 kb species of HIV-1 RNA. Reporter plasmids and identity of amplicons are indicated. At right, a summary of 3 sets of experimental results, quantitated using Image Quant v5.2 as described in (B).

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