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. 2022 Feb 2;13:630. doi: 10.1038/s41467-022-28307-7

Fig. 2. bNAbs retain mature HIV-1 particles at the plasma membrane.

Fig. 2

a Western blot analysis of Gag in CD4 T cells infected with HIV-1 (CH058) for 48 h and then cultivated 24 h with an isotype control (mGO53) or a bNAb (10–1074). α-Tubulin was used as a loading control. Molecular weights (kDa) are indicated on the left. One representative experiment (out of six) is shown. b Western blot analysis of p24 (left) and p55 (right) levels in HIV-1-infected (CH058) CD4 T cells cultivated for 24 h with an isotype control (mGO53) or a bNAb (10–1074). Results are normalized to α-Tubulin and to mGO53. Each dot represents a donor of CD4 T cells (n = 6). *p = 0.0313 (two-tailed Wilcoxon test). Bars represent the mean. c Representative confocal microscopy images of infected CD4 T cells (CH058) cultivated for 24 h with an isotype control (mGO53) or with a bNAb (10–1074). Cells were stained for intracellular total Gag (green) and p17 (red). Nuclei were stained with DAPI (blue). Scale bar: 5 μm. One representative experiment (out of five) is shown. d Confocal microscopy analysis of total Gag and p17 intensity levels in infected (CH058) CD4 T cells cultivated for 24 h with an isotype control (mGO53) or with a bNAb (10–1074). Results are expressed as the fold-change in Gag staining intensity over mGO53. Each dot represents a donor of CD4 T cells (n = 5). At least 50 cells were analyzed per donor. ***p = 0.0002 (two-tailed paired t-test). Bars represent the mean. e Representative scanning electron microscopy images of infected (CH058) CD4 T cells cultivated for 24 h with an isotype control (mGO53) or with a bNAb (10–1074). A red arrowhead points an HIV-1 viral particle as an example. Scale bar: 1 μm. One representative experiment (out of two) is shown. Antibodies were tested at a concentration of 100 nM, corresponding to 15 μg/mL. Source data are provided as a Source Data file.