Cem ss

293T, Jurkat, KG-1, Cem-A, Molt and Cem-SS cells were obtained from the American Type Culture Collection (ATCC). 293T cells were maintained in Dulbecco’s Modified Eagle Medium (Cellgro) supplemented with 10% Fetal Bovine Serum, FBS (Gemini Bioproducts). Rest of the cells were maintained in Iscove's Modified Dulbecco's Medium (ATCC) supplemented with 20% FBS.
The following reagents were obtained through the AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH; p24 Monoclonal Antibody (183-H12-5C) from Dr. Bruce Chesebro and Kathy Wehrly, pSV-Ψ-MLV-env- from Dr. Nathaniel Landau.
Goat-anti-mouse-horseradish peroxidase and goat-anti-rabbit-horseradish peroxidase (HRP) secondary antibodies and West Femto enhanced chemiluminescent (ECL) HRP substrate were obtained from Thermo Scientific (Rockford, IL). Rabbit polyclonal Antibody against gp96 was obtained from GeneTex (Irvine, CA). Mouse monoclonal antibody against GAPDH was obtained from ABM (Richmond, BC, Canada). VSVG mouse monoclonal antibody was obtained from KeraFAST. DiI-LDL was obtained from Kalen Biochemical (Montgomery Village, MD) VSV-eGFP was a kind gift from Dr. Asit Pattnaik (University of Nebraska-Lincoln). Measles-GFP was a kind gift from Dr. Stephen Russell (Mayo Clinic, Rochester).

The T-lymphoblastic leukaemia cell line, CEM-SS, (anchorage-dependent and suspension cells) were obtained from the American Type Culture Collection (ATCC), the National Cancer Institute (NCI) and the RIKEN Cell Bank (RCB).

CEM-SS and Jurkat cells were obtained from the American Type Culture Collection (ATCC). Primary CD4⁺ T cells were isolated as previously described (21 (link)). Primary CD4⁺ T cells were infected with 100 ng of WT NL4-3 or mutant viruses. Approximately 16 h later, the cells were washed and treated with dimethyl sulfoxide (DMSO) or 100 nM didehydro-cortistatin A (dCA) for 6 days. Supernatants were collected, and p24 was measured by ELISA (catalog no. 5447; Advanced BioScience Laboratories, Inc.). To obtain chronically infected Jurkat populations, cells were infected at a multiplicity of infection of 0.5 for 12 h. Cells were washed three times. Fresh culture medium was then added, and the cells were passaged twice a week for 3 months. CEM-SS and Jurkat cells cultured in Roswell Park Memorial Institute 1640 (RPMI 1640) medium with 10% fetal bovine serum (FBS) and PSG (penicillin [100 U/ml], streptomycin [100 g/ml], and l-glutamine [2 mM]). HeLa-CD4 (HT4-6C) cells were obtained from the NIH AIDS Reagent Program. Stable HeLa-CD4 LTR-Luc cells were obtained by transfecting HeLa-CD4 cells with pGL4.22_WT LTR-Luc, pGL4.22_MUT1 LTR-Luc, and pGL4.22_MUT2 LTR-Luc and selected with puromycin (2 μg/ml). HeLa-CD4 cells were cultured in Dulbecco’s modified Eagle’s medium supplemented with 5% FBS and PSG. All cells were cultured at 37°C and 5% CO₂.