Total RNA was extracted from the cells using the RNeasy Mini kit (QIAGEN) per the manufacturer's protocol. Library preparation was performed using the NEBNext Ultra II Directional RNA Library Prep Kit with rRNA Depletion Kit v2 (New England Biolabs). Library conversion was done with Singular Genomics Library Compatibility Kit (Singular Genomics). Libraries were sequenced on the Singular Genomics G4 sequencer on F3 flow cells, with one lane per library, at 75 bp pair-end for 100 cycles.