Cell Culture and Protein Expression Monitoring

HEK293T cell line was maintained in DMEM (HIMEDIA, India) and THP-1 cell line (30 (link)) in RPMI-1640 (HIMEDIA, India) media, supplemented with 10% Fetal Bovine Serum (Gibco, USA), 100 U/ml of penicillin, and 100 ug/ml of streptomycin (HIMEDIA, India) at 37°C with 5% CO₂ (31 (link)).
Different antibodies used to monitor the expression and localization of various proteins include anti-Myc antibody (Santacruz biotechnology (sc40); GAPDH (Santacruz biotechnology (sc47724); GFP antibody (Santacruz biotechnology, sc-9996 and ABclonal, AE011); anti-His antibody (Santacruz biotechnology, sc-8036), CD206 antibody (Santacruz biotechnology, sc-58986 PE), CD64 antibody (Santacruz biotechnology, sc-1184 FITC) and secondary antibodies include anti-mouse HRP (cell signaling, 08/2017), anti-rabbit HRP (Santacruz biotechnology, sc-2357) or anti-mouse Alexa 647 (Ab150115). The quantification of HIV for infection assays was performed using HIV-1 p24 Capture ELISA Assay Kit (ABL Inc, USA) and the released HIV antigens were measured using Microlisa HIV Ag & Ab assay kit (detects gp41, gp120, and p24) (JMitra &Co Pvt Ltd, India).