Fig. 2: The Ahed gene product is a nuclear protein.
From: A newly identified gene Ahed plays essential roles in murine haematopoiesis
a Confocal microscopy images of Ahedm/m ESCs expressing EGFP-tagged full length Ahed cDNA. EGFP-Ahed, Ahed cDNA tagged with EGFP at the 5ʹ-end; Ahed-EGFP, Ahed cDNA tagged with EGFP at the 3ʹ-end. Nuclei are counterstained with Hoechst 33342. Note that EGFP fluorescence is localised in the nucleus. Scale bars, 20 μm. Results shown are representative of three independent experiments. b Schematic of the AHED protein. Red boxes indicate K(K/R)X(K/R) sequences, which are distributed within the domain between a.a. residues 151–260. N, N-terminus; C, C-terminus. c Deletion of a.a. 151–260 resulted in EGFP fluorescence in both cytoplasm and nuclei (upper). Conversely, a.a. residues 151–260 are sufficient for nuclear localisation of the fusion protein (middle). Diffuse distribution of the EGFP protein is shown as a control (lower). Scale bars, 20 μm. Results shown are representative of three independent experiments. d Confocal microscopy images of Ahedm/m ESCs expressing EGFP-tagged deletion mutants (upper, deletion of a.a. 151–200; lower, deletion of a.a. 201–260). Nuclei are counterstained with Hoechst 33342. Apart from mitotic cells, EGFP fluorescence is localised in the nucleus. Red boxes in the left diagrams indicate K(K/R)X(K/R) sequences. Scale bars, 20 μm. Results shown are representative of three independent experiments.
