Journal: bioRxiv

Article Title: The primary mechanism for highly potent inhibition of HIV-1 maturation by lenacapavir

doi: 10.1101/2024.12.06.627250

Figure Lengend Snippet: (A-D) HIV-1 virions were produced by transfecting HEK293T cells (producer cells) with indicated concentrations of full-length WT pNL4.3. Indicated concentrations of LEN or DMSO control were added to HEK293T cells, and viruses isolated by ultracentrifugation through 20% sucrose cushions were used to infect HeLa TZM-bl cells (Target cells). After 48 h of infection, luciferase activity was measured to determine EC 50 values of LEN. The averaged data (+/− SD) from three independent experiments are shown.

Article Snippet: Viruses were generated by transfecting HEK293T cells (10 7 plated in 15 cm plates the previous day) with 30 µg pNL4-3 plasmid DNA using PolyJet DNA transfection reagent (SignaGen Laboratories) according to the manufacturer’s protocol.

Techniques: Produced, Control, Isolation, Infection, Luciferase, Activity Assay

Journal: bioRxiv

Article Title: The primary mechanism for highly potent inhibition of HIV-1 maturation by lenacapavir

doi: 10.1101/2024.12.06.627250

Figure Lengend Snippet: (A) The experimental design. The same virus preparation was used for three different assays: 1) LC-MS/MS to determine LEN amounts bound to virions, 2) p24 ELISA to determine CA amounts; and 3) infectivity of the viruses produced in the presence of LEN or DMSO control in HeLa TZM-bl cells. (B) Representative LC-MS/MS results of viruses produced in the presence of LEN (the top panel), the identical concentration of LEN added to HEK293T cells in the absence of pNL4.3 (the middle panel), and the control of the medium without LEN or the virus (the bottom panel). (C) Infectivity of the viruses at indicated [LEN]:[CA] ratios or DMSO control. The [LEN]:[CA] ratios were measured using p24 ELISA and LC-MS/MS results for the same virus preparations.

Article Snippet: Viruses were generated by transfecting HEK293T cells (10 7 plated in 15 cm plates the previous day) with 30 µg pNL4-3 plasmid DNA using PolyJet DNA transfection reagent (SignaGen Laboratories) according to the manufacturer’s protocol.

Techniques: Virus, Liquid Chromatography with Mass Spectroscopy, Enzyme-linked Immunosorbent Assay, Infection, Produced, Control, Concentration Assay

Journal: PLoS Pathogens

Article Title: Highly Active Antiretroviral Therapies Are Effective against HIV-1 Cell-to-Cell Transmission

doi: 10.1371/journal.ppat.1003982

Figure Lengend Snippet: ( A, B ) An experiment as in was performed for the combinations (black lines) of ( A ) AZT and TFV and ( B ) 3TC and ABC and compared to each single inhibitor treatment (red lines, left and right panel). The X-axis represents the drug concentration of the drug within the combination that is being compared to the single drug treatment (in red). See Supplementary for additional inhibitor combinations. ( C ) The change in IC 90 for co-culture over cell-free infection for the single inhibitors was compared to all the inhibitor combinations tested. ( D ) The change in average IIP at IC Max for co-culture over cell-free infection for the single inhibitors was compared to all the inhibitor combinations tested. See Supplementary for complete set of average IIP data. ( E ) Cell-free and co-culture infection of primary cells with HIV-1 NL4-3 carrying the M184V mutation of reverse transcriptase (black line) compared to wild-type HIV-1 NL4-3 (red line) in the presence of increasing concentrations of AZT. Error bars represent the standard deviation from the combination of at least two individual experiments each done in triplicate.

Article Snippet: The plasmid encoding the M184V mutation in reverse transcriptase (pNL4-3ΔEnv(M184V)) was kindly donated by Robert Siliciano, Johns Hopkins University.

Techniques: Concentration Assay, Co-Culture Assay, Infection, Mutagenesis, Reverse Transcription, Standard Deviation