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. 2019 Sep 24;10(5):e02016-19. doi: 10.1128/mBio.02016-19

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Copyright © 2019 Chao et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 6 — HEAL silencing maintains HIV-1 suppression after AZT withdrawal. (A and B) HEAL (A) and CDK2 (B) expression levels were increased in PBMCs of HIV-1-infected individuals. HEAL (A) and CDK2 (B) expression levels were measured by RT-qPCR, and signals were normalized to GAPDH mRNA levels. Data were calculated using the threshold cycle (2^−ΔΔ^CT) method and analyzed with Student’s t test. *, P < 0.05; ****, P < 0.0001. (C to E) Experimental design for the analysis of HIV-1 replication in HEAL-depleted and/or AZT-treated cells. H9 T cells were infected with control or HEAL shRNA-carrying lentiviruses, maintained under puromycin selection conditions for 3 days, and then infected with HIV-1. Cells were treated with AZT (0 to 20 μM) between days 8 and 11 and then either switched to AZT-free medium or maintained in medium containing 0 to 20 μM AZT for an additional 3 days (C). On day 14, cells were collected and analyzed for GP120 mRNA (D) or HEAL RNA (E) by RT-qPCR. Signals were normalized to GAPDH mRNA levels. n = 3, mean ± SD; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant.