4.3. Transduction of HEK-293T Cells for Transcriptomic Analysis

The day before transduction, HEK-293T cells were plated in 6 well plates (5 × 10⁵ cells/well) in DMEM supplemented with 10% FBS, 1% L-glutamine and 1% penicillin–streptomycin. The following day, the medium was discarded, and the cells were transduced with 5 ng RT-equivalent of HIV-1/HIV-2 or ”mock” pseudovirions in serum and antibiotic-free media complemented with 8 µg/mL polybrene. Cells were collected at 8, 12 and 26 h after transduction; media was discarded, and cells were washed with PBS and then suspended into TRIzol reagent (Thermo Fisher Scientific, Waltham, MA, USA). RNA isolation was carried out according to the manufacturer’s instructions. The quality of the RNA was determined by Agilent RNA 6000 Nano kit on an Agilent 2100 Bioanalyzer (Agilent Technologies, Waldbronn, Germany). Thereafter, high-throughput sequencing was performed on the MGI DNBSEQ G400 sequencer using MGIEasy RNA Library Prep Set at the Genomic Medicine and Bioinformatics Core Facility of the University of Debrecen.