Journal: bioRxiv

Article Title: M-Sec promotes the production of infectious HIV-1 virus through the exocyst complex

doi: 10.1101/2025.11.13.688207

Figure Lengend Snippet: ( A ) The control or stable M-Sec knockdown (ΔM-Sec) U87 cells were left uninfected or infected with HIV-1, cultured for 2 days, and analyzed for their expression level of Env (gp160 and gp120 in the top blot, and gp41 in the middle blot) by western blotting. β-actin blot is the loading control. ( B ) The control or ΔM-Sec U87 cells were left uninfected or infected with HIV-1, and cultured for 2 days. The virus-like particles in the supernatants were collected by centrifugation, and analyzed for their amount of gp120 (top), gp41 (middle), and p24 Gag (bottom) by western blotting. ( C ) The virus-like particles were analyzed as in B . The density of gp120- or gp41 band was normalized to that of p24 Gag band, and represented as a percentage relative to that of the control cells (n=3). * p < 0.05. ( D , E ) The control or ΔM-Sec U87 cells were infected with HIV-1, and cultured for 2 days. In D , the supernatants were collected, and analyzed for the activity of reverse transcriptase (RT) by qPCR (n=4). In E , the supernatants were collected, and analyzed for viral infectivity using TZM-bl cells as the target cells. Two different inputs were tested (2 and 4 nU/mL of RT activity). The infectivity is represented as a percentage relative to that of the control cells with the input of 2 nU/mL (n=3).

Article Snippet: The cells were fixed with paraformaldehyde, permeabilized with Triton X-100, and stained with anti-p24 Gag (24-2; Santa Cruz Biotechnology) or anti-gp120 Env (2G12; Syd Labs) followed by AlexaFluor633-labeled anti-mouse IgG or AlexaFluor568-labeled anti-human IgG (both from Molecular Probes).

Techniques: Control, Knockdown, Infection, Cell Culture, Expressing, Western Blot, Virus, Centrifugation, Activity Assay, Reverse Transcription, Quantitative RT-PCR

Journal: bioRxiv

Article Title: M-Sec promotes the production of infectious HIV-1 virus through the exocyst complex

doi: 10.1101/2025.11.13.688207

Figure Lengend Snippet: ( A ) The parental or stable M-Sec-expressing 293 cells were transfected with the empty plasmid or HIV-1 molecular clone, cultured for 2 days, and analyzed for their expression level of gp160 by western blotting. β-actin blot is the loading control. ( B ) The parental or M-Sec-expressing 293 cells were transfected with the empty plasmid or HIV-1 molecular clone, and cultured for 2 days. The virus-like particles in the supernatants were collected by centrifugation, and analyzed for their amount of gp120 (top), gp41 (middle), and p24 Gag (bottom) by western blotting. ( C ) The cells were analyzed as in B . The density of gp120- or gp41 band was normalized to that of p24 Gag band, and represented as a relative to that of the parental cells (n=3). * p < 0.05. ( D , E ) The parental or M-Sec-expressing 293 cells were transfected with the HIV-1 molecular clone, and cultured for 2 days. In D , the supernatants were collected, and analyzed for the activity of reverse transcriptase (RT) by qPCR (n=5). In E , the supernatants were collected, and analyzed for viral infectivity using TZM-bl cells as the target cells. Two different inputs were tested (2 and 4 nU/mL of RT activity). The infectivity is represented as a percentage relative to that of the parental cells with the input of 2 nU/mL (n=3).

Article Snippet: The cells were fixed with paraformaldehyde, permeabilized with Triton X-100, and stained with anti-p24 Gag (24-2; Santa Cruz Biotechnology) or anti-gp120 Env (2G12; Syd Labs) followed by AlexaFluor633-labeled anti-mouse IgG or AlexaFluor568-labeled anti-human IgG (both from Molecular Probes).

Techniques: Expressing, Transfection, Plasmid Preparation, Cell Culture, Western Blot, Control, Virus, Centrifugation, Activity Assay, Reverse Transcription, Quantitative RT-PCR, Infection