human t lymphoblast cell line sup t1  (ATCC)

Journal: bioRxiv

Article Title: The stage-specific regulation imposed by Importinβ-1 on HIV-1 propagation and infectivity dynamics

doi: 10.1101/2024.06.10.598175

Figure Lengend Snippet: (A) Immunoblot showing the presence of Importinβ-1 in virus preparation from culture supernatants of pNL4.3 transfected HEK293T cells. The presence of virus was detected with anti-p24 antibody. GAPDH was used as the internal control. ( B) Immunoblot showing the presence of Importinβ-1 in virus preparation from culture supernatants of NL4.3 infected SUP-T1 cells. The presence of virus was detected with anti-p24 antibody. GAPDH was used as the internal control. (C) Immunoblot showing the presence of Importinβ-1 in virus preparation from culture supernatants of NL4.3 infected 1321N1 cells. The presence of virus was detected with anti-p24 antibody. GAPDH was used as the internal control. (D) Co-immune precipitation showing Importinβ-1 interaction with Gag and Capsid proteins upon pNL4.3 transfection in HEK293T cells. Mouse IgG was used as an isotypic control. (E) Confocal microscopy showing co-localisation of Importinβ-1 GFP and Gag/Capsid protein in the HEK293T cells. (F) The plot profiles showing the overlapping of Importinβ-1 GFP and Gag/Capsid protein in representative single cell generated using ImageJ software. All experiments were done at least 3 times.

Article Snippet: DMEM (HIMEDIA Cat: AL066A-500ML) was used for maintaining HEK293T, 1321N1, and TZM-bl cell lines, and RPMI-1640 (HIMEDIA Cat: AL162A-500ML) for SUP-T1 cell line.

Techniques: Western Blot, Virus, Transfection, Infection, Confocal Microscopy, Generated, Software

Journal: bioRxiv

Article Title: The stage-specific regulation imposed by Importinβ-1 on HIV-1 propagation and infectivity dynamics

doi: 10.1101/2024.06.10.598175

Figure Lengend Snippet: (A) Representative Immunoblot showing the levels of Importinβ-1 from 0hrs to 96hrs in SUP-T1 cells upon NL4.3 infection. GAPDH was used as a control. (B) Bar graph representing the quantification of Importinβ-1 for the experiments represented in A. (C) Levels of released virus determined by p24 ELISA in the supernatant of NL4.3 infected SUP-T1 cells from 0 hrs to 96 hrs. (D) Line graph representing the relationship of endogenous Importinβ-1 and HIV titers in SUP-T1 cells during the course of HIV infection from 0 hrs to 96 hrs. Endogenous GAPDH was used as a loading control. (E) Representative Immunoblot showing the levels of Importinβ-1 in SUP-T1 cells upon treatment with IFNγ, LPS, and IL4. Tubulin was used as a control. Bar graph showing immunoblot quantification of Importinβ-1 from three independent experiments. (F) Representative Immunoblot showing the levels of Importinβ-1 in HEK293T cells upon treatment with IFNγ, LPS, and IL4. Tubulin was used as a control. Bar graph showing immunoblot quantification of Importinβ-1 from three independent experiments. (G) Representative Immunoblot showing the levels of p55 in SUP-T1 cells upon treatment with IFNγ, LPS, and IL4. GAPDH was used as a control. Bar graph showing immunoblot quantification of Importinβ-1 from three independent experiments. All experiments were done at least 3 times. The significance is determined using an unpaired student’s t-test. The p values are denoted as *** p <0.001; * p <0.05, while non-significant values are denoted by n.s.

Article Snippet: DMEM (HIMEDIA Cat: AL066A-500ML) was used for maintaining HEK293T, 1321N1, and TZM-bl cell lines, and RPMI-1640 (HIMEDIA Cat: AL162A-500ML) for SUP-T1 cell line.

Techniques: Western Blot, Infection, Virus, Enzyme-linked Immunosorbent Assay