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. 2014 Dec 8;43(1):336–347. doi: 10.1093/nar/gku1232

Figure 4.

Figure 4.

Late assembly of ΔZF2 HIV-1 and viral production were trans-complemented by targeting Tsg101 to the PM (A) trans-complementation strategy. A chimeric ΔZF2Gagc protein was used to target Tsg101 to the PM (ΔZF2Gagc-Tsg101) and to restore ΔZF2 virus biogenesis (left panel). Similar experiments were conducted with WT proteins (right panel). A protocol similar to the one used in Figure 1 was used to transfect HeLa cells with the two pairs of proviral plasmids and trans-complementation was performed with a third pENX vector series encoding chimeric Gagc proteins deleted of p6 (Text S7). (B) Cells were observed by TIRFM 14 h post-transfection and the area of a thousand individual virus puncta was determined. Data are presented as a scatter plot of the areas of individual puncta. ***P < 0.001 using the Mann–Whitney U-test. (C) Virus production in trans-complementation experiments. Virus-containing supernatants of transfected cells were collected and the amounts of viruses were determined by p24 ELISA. Values were normalized to the level of WT particles measured in the context of trans-complementation (wtHIV + wtGagc). **P < 0.01 using the Student's t-test.