GSM9463701: HFF-1 cells,uninfected HSV-2,24h,Control2; Homo sapiens; OTHER
HFF-1 cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM, Vivacell, cat. no. C3113-0500) containing 20% fetal bovine serum (FBS; Cell-Box, cat. no. AU3-01S-02-S) and penicillin-streptomycin (100 U/mL) at 37°C under 5% CO₂. HFF cells were seeded into T225 flasks and cultured until reaching approximately 90% confluence after 24 hours. The cells were then washed twice with phosphate-buffered saline (PBS), and three flasks were infected with HSV-2 G at an MOI of 0.1 to form the infection group, while three other flasks remained uninfected as negative controls. After adsorption for 1 hour at 37°C, fresh DMEM supplemented with 20% FBS was added, and the cells were further cultured for 24 hours before harvesting the cell samples. following the manufacturer's instructions. Briefly, RNA was randomly fragmented to ~200 nt by RNA Fragmentation Reagents. Protein A/G beads were coupled to the m6A antibody by rotating at room temperature for 1 h. The RNA fragments were incubated with the bead-linked antibodies and rotated at 4°C for 4 h. After incubation, the RNA/antibody complexes were washed for several times, and then, captured RNA was eluted from the complexes and purified. RNA libraries for IP and input samples were then constructed with GenSeq® Low Input Whole RNA Library Prep Kit(GenSeq, Inc.) by following the manufacturer's instructions. Libraries were qualified using Agilent 2100 bioanalyzer (Agilent) and then sequenced in a DNBSEQT7 platform (MGI Tech).