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. 2017 Jul 28;6:e26067. doi: 10.7554/eLife.26067

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© 2017, Snapp et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 8. — (A) SupT1 cells (5x10⁶⁾ were infected with 20 µg of wild-type or M26P-mutant pLAI DNA constructs using electroporation. Replication was monitored by CA-p24 ELISA. The replication curves shown represent the averaged values of 6 independent experiments. p=0.0058 (**) and 0.003 (***). (B) SupT1 cells (Krogh et al., 2001) were infected with a total of 50 pg CA-p24 equivalent wild-type or M26P-mutant LAI virus (produced by transfected HEK293T cells) in ratios 1:1 and 1:10, each in duplicate (biological replicate). After 4 and 35 days, the isolated virus from each culture was sequenced and the electropherograms were quantified. (C,D,E) TZM-bl reporter cells were infected with: 500 pg of wild-type or M26P LAI virus (produced by HEK293T cells) (n.s, p=0.0833) (C), 1,000 pg of wild-type or M26P-mutant JR-FL pseudo-virus (produced by HEK293T cells) (p<0.0001). One M26P data point was excluded as it lay well below the background. (D), 1,000 pg of wild-type, Ig κ (p=0.0004) or cystatin-SP (p=0.0005) mutant LAI pseudo-virus (produced by C33A cells) (E) and infectivity was measured by Luciferase activity. The data in panels A, C and D are derived from 3 independent experiments, using 3 independently produced (pseudo-)virus stocks (biological replicates), each performed in quadruplicate (technical replicates). The data in panel B are derived from 2 independent experiments, using 2 independently produced virus stocks (biological replicates) and sequenced with two independent primers (technical replicates). The data in panel E are derived from 1 pseudovirus stock (biological replicate), performed in quadruplicate (technical replicates). All significance values were calculated with an unpaired, two-tailed t test with Welch’s correction.

Figure 8—figure supplement 1. — (A) SupT1 cells (5x10⁶⁾ were infected with 20 µg of wild-type or M26P-mutant pLAI DNA constructs using electroporation. Replication was monitored by CA-p24 ELISA. The replication curves shown represent the averaged values of 6 independent experiments. p=0.0058 (**) and 0.003 (***). (B) SupT1 cells (Krogh et al., 2001) were infected with a total of 50 pg CA-p24 equivalent wild-type or M26P-mutant LAI virus (produced by transfected HEK293T cells) in ratios 1:1 and 1:10, each in duplicate (biological replicate). After 4 and 35 days, the isolated virus from each culture was sequenced and the electropherograms were quantified. (C,D,E) TZM-bl reporter cells were infected with: 500 pg of wild-type or M26P LAI virus (produced by HEK293T cells) (n.s, p=0.0833) (C), 1,000 pg of wild-type or M26P-mutant JR-FL pseudo-virus (produced by HEK293T cells) (p<0.0001). One M26P data point was excluded as it lay well below the background. (D), 1,000 pg of wild-type, Ig κ (p=0.0004) or cystatin-SP (p=0.0005) mutant LAI pseudo-virus (produced by C33A cells) (E) and infectivity was measured by Luciferase activity. The data in panels A, C and D are derived from 3 independent experiments, using 3 independently produced (pseudo-)virus stocks (biological replicates), each performed in quadruplicate (technical replicates). The data in panel B are derived from 2 independent experiments, using 2 independently produced virus stocks (biological replicates) and sequenced with two independent primers (technical replicates). The data in panel E are derived from 1 pseudovirus stock (biological replicate), performed in quadruplicate (technical replicates). All significance values were calculated with an unpaired, two-tailed t test with Welch’s correction.