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. 2009 Oct 5;284(48):33384–33391. doi: 10.1074/jbc.M109.021659

FIGURE 4.

FIGURE 4.

Influences of the expression levels of hnRNP A1, Q, R, K, and U on HIV production by persistently infected astrocytes. A, overexpression of the hnRNP A1 protein increased Gag p24 production by TH4-7-5 cells. TH4-7-5 cells were transfected with plasmids for expression of hnRNP A1 fused to a blue fluorescent tag (hnRNP A1-CYN) or with plasmids for expression of the unlinked blue fluorescent protein (CYN) as control. Gag production was monitored by measuring intra- and extracellular levels of Gag p24. Gag p24 levels are indicated relative to Gag levels in cells transfected with the CYN control plasmid. Expression of ectopic proteins was confirmed by Western blot analysis. B, siRNA-mediated knock-down of endogenous hnRNP A1 expression reduced Gag p24 produced by TH4-7-5 cells. Gag p24 levels are indicated relative to Gag levels in cells transfected with non-silencing (n.s.) siRNAs. Western blot analysis confirms knockdown of hnRNP A1. C, investigation of the effects of hnRNP Q, R, K, and U knock-down on HIV production by TH4-7-5 cells. Knock-down of hnRNP Q, R, and K reduced Gag p24 production by TH4-7-5 cells. Knock-down of hnRNP U slightly increased Gag production. Gag p24 levels are indicated relative to Gag levels in cells transfected with non-silencing siRNAs. Western blot analysis confirms knock-down of each hnRNP.