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Figure 3 | Retrovirology

Figure 3

From: A novel HIV-1-encoded microRNA enhances its viral replication by targeting the TATA box region

Figure 3

Effect of miR-H3 on HIV-1 viral replication. (A) Ectopic expression of miR-H3 by a construct containing its precursor. Mature miR-H3-3p sequence was tested with Real-time qPCR, an empty vector was transfected as control. (B) HEK293T cells were co-transfected with an Env-defective HIV-1 clone named pNL4-3-deltaE-EGFP and a miR-H3 precursor (pre-miR-H3) or the empty vector pEGFP-C1. The productions of P24 in the supernatant were determined at 48 hrs post-transfection. (C) HEK293T cells were transfected with pNL4-3-deltaE-EGFP or a miR-H3 mutant named pNL4-3-deltaE-EGFP miR-H3-mut. The productions of P24 in the supernatant were determined as described above. (D) Silent mutations were introduced to miR-H3 precursor (labeled in red with underline) to eliminate its processing without altering the amino acids of the HIV-1 RT gene (top). The reverse transcriptase activity of wildtype or miR-H3 deficient viruses was tested at different concentrations (bottom). (E) Equal amount (5 ng of P24 antigen) of wildtype and mutated viruses were used for the infection of 2x106 activated primary CD4+ T-lymphocytes. The viral production in the supernatant was assayed by P24 ELISA at several time intervals. P-values were calculated using the two tailed unpaired Student’s t-test with equal variances, n = 3. *p < 0.05, **p < 0.01.

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