Fig. 4: NCOR2-013 suppresses activation of JunB by forming a transcription-regulatory complex with HDAC3 and TBL1XR1/TBLR1.

a–g The human NCOR2-013 overexpressing THP-1 macrophages were infected with T. marneffei conidia at MOI of 10 for 24 h. a The flowchart of potential binding protein screen for NCOR2-013. Briefly, anti-Flag antibody (for Flag-NCOR2-013) and isotype control IgG were used for immunoprecipitation of proteins, respectively, and LC-MS/MS was used to detect proteins that bound to NCOR2-013. The NCOR2-013 binding proteins were then intersected with DEGs (T. marneffei-infected NCRO2-013 overexpressing THP-1 macrophages vs. T. marneffei-infected control cells) and TFs (list of human transcription factors). b IP was performed with anti-Flag antibody (for Flag-NCOR2-013), and WB was used to detect NCOR2/SMRT, TBL1XR1/TBLR1, HDAC3 and JunB expression. IP assays were performed with JunB (c), TBL1XR1 / TBLR1 (d), and HDAC3 (e) antibody, respectively, to show interactions between NCOR2-013 and TBL1XR1/TBLR1, HDAC3, JunB, respectively. Co-IP assays show interactions between Flag-NCOR2-013 and JunD (f) or c-Jun (g). NCRO2-013 overexpressing THP-1 macrophages and control cells were infected with T. marneffei for 24 h and 48 h, p-JunB and JunB (h), p-c-Jun and c-Jun (i) were detected by WB. β-actin was used as the loading control. Experiment was performed thrice (n = 3 biological replicates) and a representative blot is shown. Supplementary material is available (Supplementary Figs. 2, 6). MOI multiplicity of infection, DEGs differential expression genes, TFs transcription factors, NCOR2-013 OE NCRO2-013 overexpression, Tm T. marneffei.