PHLPP1 Cut&Tag profiling of bone marrow derived macrophage

Purpose: To transcriptome widely reveal genes regulation by PHLPP1, we performed Cleavage Under Targets and Tagmentation (CUT&Tag) analysis of wild-type (WT) BMDMs. Methods: Mouse BMDMs were generated from bone marrow cells in RPMI-1640 medium with recombinant mouse M-CSF (20ng/ml). BMDMs were stained to confirm the surface expression of CD11b and F4/80. Cells with purity >97.5% were used for subsequent experiments. WT BMDMs were stimulated with IL-4 (20ng/ml) for 4 hours. Results: We carried out a CUT&Tag assay using antibodies against PHLPP1, and found the global influence of PHLPP1 in regulating genes of transcription regulatory regions of key genes for pro-fibrosis macrophage effector function. Overall design: Cleavage Under Targets and Tagmentation (CUT&Tag) analysis for PHLPP1-binding DNA in WT BMDMs stimulated with IL-4.