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 and stable M-Sec knockdown (ΔM-Sec) U87 cells were analyzed for their expression level of M-Sec by western blotting. β-actin blot is the loading control. ( B ) The control or ΔM-Sec U87 cells were infected with HIV-1, cultured for 2 days, and analyzed for Gag (red) by immunofluorescence. The nuclei were stained with DAPI (blue). In the right panels, the magnified images of "a" and "b" in the left panels are shown. Scale bars: 40 μm and 10 μm for the left and right panels, respectively. ( C ) The cells were analyzed as in B . In the upper panel, the average size of Gag puncta in each Gag + cell is summarized (15 cells for each group). In the lower panel, the number of >0.25 μm Gag puncta in each Gag + cell is summarized (15 cells for each group). The Gag signal larger than approximately 0.03 μm was considered puncta. * p < 0.05.

Article Snippet: The U87 (U-87MG) glioma cell line was obtained through the American Type Culture Collection (ATCC) and maintained with DMEM-10% FCS.

Techniques: Control, Knockdown, Expressing, Western Blot, Infection, Cell Culture, Immunofluorescence, Staining

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 infected with HIV-1, cultured for 2 days, and analyzed for Env (green) and Gag (red) by immunofluorescence. The nuclei were stained with DAPI (blue). In the right panels, the magnified images of "a" and "b" in the middle panels are shown. Scale bars: 10 μm and 5 μm for the left/middle and right panels, respectively. ( B ) The cells were analyzed as in A . The co-localization of Gag and Env was quantified as Pearson’s correlation coefficients (PCCs) (10 cells for each group). * p < 0.05.

Article Snippet: The U87 (U-87MG) glioma cell line was obtained through the American Type Culture Collection (ATCC) and maintained with DMEM-10% FCS.

Techniques: Control, Knockdown, Infection, Cell Culture, Immunofluorescence, Staining

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 U87 (U-87MG) glioma cell line was obtained through the American Type Culture Collection (ATCC) and maintained with DMEM-10% FCS.

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 - C ) The control U87 cells were pretreated with DMSO, 10 μM BQU57 or 10 μM ES2 for 2 days, and infected with HIV-1. Then, the cells were cultured for 2 days in the presence of DMSO, 10 μM BQU57 or 10 μM ES2, and analyzed for Env (green) and Gag (red) by immunofluorescence. The nuclei were stained with DAPI (blue). Scale bar: 10 μm. In the upper panel of B , the average size of Gag puncta in each Gag + cell is summarized (10 cells for each group). In the lower panel of B , the number of >0.25 μm Gag puncta in each Gag + cell is summarized (10 cells for each group). The Gag signal larger than approximately 0.03 μm was considered puncta. In C , the co-localization of Gag and Env was quantified as Pearson’s correlation coefficients (PCCs) (10 cells for each group). * p < 0.05.

Article Snippet: The U87 (U-87MG) glioma cell line was obtained through the American Type Culture Collection (ATCC) and maintained with DMEM-10% FCS.

Techniques: Control, Infection, Cell Culture, Immunofluorescence, Staining