Figure 3

HnRNP Q overexpression elevates Gag-p24 production without affecting levels of HIV-1 transcripts. A) Chronically HIV-1 infected HeLa cells (LC5-HIV) were used to evaluate the effects of hnRNP Q overexpression on HIV-1 replication. Using ELISA, p24 protein levels were analyzed in LC5-HIV cells ectopically expressing hnRNP Q-CYN or CYN. The results revealed that both extra- and intracellular p24 protein levels are elevated by hnRNP Q overexpression. B) Specific primers (indicated in the scheme of the HIV-1 genome) were designed to distinguish between Rev-dependent late transcripts (i.e. unspliced and singly-spliced), and Rev-independent early transcripts (i.e. completely spliced) and all transcripts. The primers for detection of late transcripts were placed into the gag ORF and for detection of all transcripts into the nef ORF. We used a 5′ spanning primer bridging the two rev exons and a 3′ primer located at the end of the second rev exon to quantify early transcripts by qRT-PCR. C) Quantitative analyses of HIV transcripts showed that levels of neither early nor late transcripts are affected by ectopic expression of hnRNP Q in chronically HIV-1 infected cells. D) Analysis of total levels of HIV-1 transcripts confirmed that overexpression of hnRNP Q did not affect HIV-1 transcript levels. p-values: * = p ≤ 0.05; ** = p ≤ 0.01; n.s. = not significant (p > 0.05).