Bright glotm luciferase assay system kit

For HIV-1 infection assay, sorted TZM-bl cells (1 × 10⁵) were seeded in a 24-well plate one day before HIV-1 infection. These cells were then challenged with HIV-1_NL4-3, HIV-1_BH10, HIV-1_HXB2 or HIV-1_YU2 for 8 hours in a total volume of 500 µl serum-free medium. After removing the culture medium and washing cells with PBS 3 times, the cells were cultured in fresh complete media for indicated days. Then, the cells were harvested for luciferase activity analysis with a Bright-GloTM Luciferase Assay System Kit (Promega, WI, USA) by a luminometer (Promega Glomax, USA). At the same time, the supernatant was collected for HIV-1 p24 antigen detection using an ELISA kit (ZeptoMetrix).

A plasmid‐based reverse genetics system was used to evaluate influenza viral RNA polymerase activity using Bright‐GloTM Luciferase Assay System kit (E2610, Promega) as previously described.²⁰, ²¹ With Lipofectamine™ 3000 Transfection Reagent (Thermo Fisher), 293T cells were transfected with plasmids expressing IAV (A/Udorn/1972) PB1, PB2, PA, NP, and pHH21‐Luc for 6 h and were subsequently treated with Phillyrin, FS21, or ribavirin (as positive control) for 24 h. Cell lysates were then harvested, and luciferase activity was measured using Wallac Victor (Perkin Elmer) according to manufacturer's instructions.