Journal: bioRxiv

Article Title: Cryoelectron tomography of HIV-1 cell-cell transmission conjugates reveals a secluded environment for viral assembly and transfer

doi: 10.64898/2026.06.11.725753

Figure Lengend Snippet: ( A ) Comparison NL4-3 Envelope and Δ Env sequences. Dotted box indicates sequence deletion and hashed box with red cross indicates lack of translation due to frameshift and introduction of a stop codon ( B ) The percentage of all (single-cell and conjugate) GFP+CTDR+ events at 3h or 24h co-cultures of uninfected SupT1 R5 cells mixed with either NL4-3 IRES-eGFP or Δ Env IRES-eGFP infected SupT1 R5 cells. ( C ) Representative imaging cytometry of single cells and conjugates present in NL4-3 IRES-eGFP and Δ Env IRES-eGFP co-cultures at 3h and 24h. ( D ) Breakdown of the distribution of single cell and conjugate GFP+CTDR+ events within the 3-hour or 24-hour co-cultures. Data from N=1 experiment that was performed in duplicate.

Article Snippet: After 3 hours, co-cultures were fixed with 4% PFA for 30 minutes at room temperature and permeabilized with saponin 0.1% (ThermoFisher, #J63209.AK) for 10 minutes, followed by a blocking step with 2% normal goat serum and 1% BSA for 15 minutes.

Techniques: Comparison, Sequencing, Single Cell, Infection, Imaging, Cytometry

Journal: bioRxiv

Article Title: Cryoelectron tomography of HIV-1 cell-cell transmission conjugates reveals a secluded environment for viral assembly and transfer

doi: 10.64898/2026.06.11.725753

Figure Lengend Snippet: ( A ) Representative dot plots of multicell conjugates in 3h co-cultures between target CTDR+ SupT1 R5 cells mixed with either NL4-3 IRES-eGFP or ΔEnv IRES-eGFP infected SupT1 R5 cells. ( B ) The percentage of GFP+CTDR+ multicell conjugate events from initial 0-hour or 3-hour co-cultures. ( C ) Representative dot plots of single cells in 24-hour co-cultures of target CTDR+ SupT1 R5 cells mixed with either NL4-3 IRES-eGFP or ΔEnv IRES-eGFP+ infected SupT1 R5 cells ( D ) The percentage of GFP+CTDR+ single cell events from initial 0-hour or 24-hour co-cultures. Distinct shapes in B and D each represent distinct biological replicates (N = 3) and each individual shape a technical replicate. (E-H) Same as A-D but with co-culture of infected GFP+ primary CD4+ T lymphocytes and target CTDR+ primary CD4+ T lymphocytes. Shapes in F and H represent distinct primary cell donors (N = 4). All values are normalized to initial GFP+ infection levels prior to co-culture. Values below x-axis indicate the mean and standard deviation for each category. Black bar represents the mean and the gray box represents ±1 standard deviation. P-values calculated using either two-tailed unpaired t- test ( B and D ) or two-tailed paired t- test ( F and H ).

Article Snippet: After 3 hours, co-cultures were fixed with 4% PFA for 30 minutes at room temperature and permeabilized with saponin 0.1% (ThermoFisher, #J63209.AK) for 10 minutes, followed by a blocking step with 2% normal goat serum and 1% BSA for 15 minutes.

Techniques: Infection, Single Cell, Co-Culture Assay, Standard Deviation, Two Tailed Test

Journal: bioRxiv

Article Title: Cryoelectron tomography of HIV-1 cell-cell transmission conjugates reveals a secluded environment for viral assembly and transfer

doi: 10.64898/2026.06.11.725753

Figure Lengend Snippet: ( A , D ) Quantification of GFP+CTDR+ multicell conjugates events in 3-hour co-cultures of target CTDR+ SupT1 R5 and NL4-3 IRES-eGFP infected SupT1 R5 cells treated with varying concentrations of soluble CD4 (sCD4, A ) or anti-CD4 (SIM.2, D ). ( B and E ) Quantification of GFP+CTDR+ single cells in 24-hour co-cultures treated with varying concentrations of sCD4 ( B ) or SIM.2 ( E ). ( C and F ) Quantification of GFP+ SupT1 R5 after cell-free NL4-3 IRES-eGFP virus infection treated with varying concentrations of sCD4 ( C ) or SIM.2 ( F ). Data are derived from distinct biological replicates (N = 2), where colors represent distinct biological and each dot a technical replicate. All values are normalized to untreated controls and initial GFP+ infection levels prior to co-culture (if applicable). Values below x-axis indicate the mean and standard deviation for each category. Black bar represents the mean and the gray box represents ±1 standard deviation. P-values calculated using one-way ANOVA with a Tukey’s test for multiple comparisons.

Article Snippet: After 3 hours, co-cultures were fixed with 4% PFA for 30 minutes at room temperature and permeabilized with saponin 0.1% (ThermoFisher, #J63209.AK) for 10 minutes, followed by a blocking step with 2% normal goat serum and 1% BSA for 15 minutes.

Techniques: Infection, Virus, Derivative Assay, Co-Culture Assay, Standard Deviation