Homo sapiens
Protocols: Cells were treated with 2 μg/ml doxycycline and 0.06 μg/ml nocodazole, 24 and 18 h respectively, before the addition of 1 mM IAA to induce degradation of mAID-tagged proteins. After 4 h treatment with IAA, mitotic cells were manually detached by shake-off, and cells were transferred to new poly-L-lysin coated dishes. Once attached, cells were washed 3x with PBS and released from mitosis into G1 for 18 h in a fresh medium containing 2 mM thymidine.Cells were washed with PBS, detached with TrypLE and pelleted by centrifugation. Total RNA was extracted from HCT116 cells (G1, -/+ mAID-protein of interest) using Monarch Total RNA Miniprep Kit (New England Biolabs), according to manufacturer's instructions. Total RNA were sent in dry-ice package to Macrogen Europe B.V. Macrogen Europe BV constructed libraries using Illumina TruSeq stranded mRNA library preparation with Ribozero rRNA depletion.