The application of LIMe-ID to study the impact of PRC2 perturbations on lamina assoication.

We developed and performed LIMe-ID to simultaneously measure lamina association and DNA methylation in one experimental workflow. We then conducted LIMe-ID following perturbations of PRC2 to better understand the influence of H3K27me3 on lamina association. LIMe-ID experiments were performed in duplicate for either the M.CviPI-Lamin-B1 K562 cell line following inhibitor/vehicle treatments or the FLAG-NLS-M.CviPI-Lamin-B1 K562 EZH2 knock-in cell line following dTAG-13/vehicle treatment. EZH2 inihibtion, EED inhibition and vehicle (DMSO) treatment were each performed in duplicate for 72 hours with doxycycline added for the final 24 hours of the drug treatment for the inhibitor LIMe-ID. For the degradation LIMe-ID experiment, dTAG-13 and vehicle (DMSO) treatment were each performed in duplicate for 72 hours with doxycycline added for the final 24 hours of the drug treatment.