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Figure 4 | Retrovirology

Figure 4

From: Inhibition of HIV-1 gene expression by Ciclopirox and Deferiprone, drugs that prevent hypusination of eukaryotic initiation factor 5A

Figure 4

Inhibition of HIV RNA expression from molecular clones. A. Schematic of HIV-1 provirus showing major transcripts, the position of the antisense probe, and fragments protected by RPA from spliced (S) and unspliced (U) transcripts. The positions of the Rev start codon mutation in pMRev(-) and the FF substitution in pNL4-3-LucE- are marked with one and two asterisks, respectively. B. Cytoplasmic and nuclear RNA isolated at 12 hr from 293T cells co-transfected with pNL4-3-LucE- and pCMV-Ren. Drugs were added where indicated at concentrations specified in Fig. 2D. RNA was isolated at 12 hr post-transfection. Autoradiograms display RPA fragments corresponding to HIV and Renilla RNAs (upper and middle panels, respectively). Renilla RNA was analyzed as in Fig. 3. The lower panel displays quantitation of protected spliced and unspliced RNA fragments relative to the Renilla RNA fragment (mean of 2 experiments in duplicate, ± SD). Probe: undigested probe in an amount equivalent to 5% of the input to the protection assays was run as a control. C. Effect of Rev. RNA from 293T cells transfected with the Rev-defective HIV molecular clone pMRev(-) together with (+) or without (-) Rev expression vector. RNA was isolated at 15 hr post-transfection. The lower panel displays quantitation of protected spliced and unspliced RNA fragments relative to the cytoplasmic unspliced control RNA (mean of 2 experiments in duplicate, ± SD).

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