library_construction_protocol: Human primary total T cells were isolated from the blood sample of three donors from Gulf Coast Regional Blood Center using Ficoll density gradient method. The cells were collected and washed with PBS + 0.5% FBS and taken into culture. After isolation, the cells were activated with anti-CD3/CD28 microbeads (Thermo Fisher) for 24 hours. For transcription inhibition, the cultured cells were treated with ActD (10 μg/ml), for four time points, 0 hour (immediately after ActD treatment), 15 minutes, 1 hour and 2 hours. After treatment, RNA was isolated by RNeasy kit (Qiagen). Samples were sent to BGI for library preparation and follow-up sequencing