MYRF Is a Membrane-Associated Transcription Factor That Autoproteolytically Cleaves to Directly Activate Myelin Genes
(A) Schematic of the MYRF protein showing positions of the predicted NLS, Ndt80-like DBD, ICD, coiled coil region, and transmembrane region. (B) Western blot analysis of a double tagged (N-terminal Myc, C-terminal FLAG tagged) or untagged MYRF expression construct in 293T cells. Probing with anti-Myc or anti-FLAG reveals the presence of a ∼140 kDa full-length fragment and truncated cleavage products. (C) Western blot of control and Myrf CKO cultured mouse oligodendrocyte lysates with anti-C-terminal MYRF mAb. (D) Cell fractionation experiment showing that the majority of Myc-tagged N-terminal cleaved MYRF product is present in the nucleus of 293T cells (Anti-Hsp60 and anti-Parp used for cytoplasmic and nuclear controls, respectively). (E) CG-4 cells transfected with the Myc-MYRF-FLAG construct were co-stained for anti-FLAG and either anti-Myc, anti-Calnexin, or anti-golgi matrix protein 130 (gm130). (F–H) Adult mouse optic nerve stained with the anti-N-terminal-MYRF and anti-C-terminal-MYRF-mab antibodies (F), anti-C-terminal-MYRF-mab and anti-Sox10 (G), or anti-N-terminal-MYRF-mab and CC1 (H).