Characterization of the GTPase family member Gimap4. (A) In vitro transcription/translation of Gimap4 cDNA revealed a 38 kDa protein. Positive control (PC) (51 kDa luciferase) and negative control (NC) (the empty vector) are included. (B) A motif search revealed a P loop (bold, italic), an IQ domain (bold), and several PKC phosphorylation sites (bold, underlined). (C) The protein expression pattern of Gimap4 was determined by Western blotting on sorted DN, DP, and SP thymocytes using polyclonal rabbit antibodies directed against various parts of mouse Gimap4. Blots were reprobed with anti–β-actin as a loading control. (D) Western blot of sorted hematopoietic subsets: B cells (B), CD4⁺ T cells, CD8⁺ T cells, natural killer cells (NK), macrophages (MP), and dendritic cells (DC). Thymocytes from a Rag ko mouse and splenocytes from Gimap4 ko (see Figure 5) served as a negative control; splenocytes from a wild-type (wt) mouse, as a positive control.