Fig. 4

NUP98-NSD1 retains nuclear localization pattern and interactions in murine NUP98-NSD1/FLT3-ITD immortalized hematopoietic stem and progenitor cells and in NUP98-NSD1⁺ patient cells. A Immunofluorescence staining showing the nuclear puncta pattern of localization of the fusion in primary murine NUP98-NSD1/FLT3-ITD immortalized hematopoietic cells. The cells were stained with Anti-FLAG antibody labeled with Alexa Fluor 568 secondary antibody (green). Nuclei were counterstained with DAPI (blue). B Proximity ligation assay in NUP98-NSD1/FLT3-ITD immortalized hematopoietic cells showing the interaction between NUP98-NSD1 and SMARCA5/BPTF, detected using kit specific fluorophore (lem = 669 nm, far red). Nuclei were counterstained with DAPI (blue). C Immunofluorescence staining showing the nuclear puncta pattern of localization of the fusion in NUP98-NSD1⁺ patient cells. The cells were stained with Anti-NSD1 and Anti-NUP98 antibodies targeting the C-terminus and N-terminus (epitopes included in the fusion protein), respectively. Primary antibodies were then labeled with Alexa Fluor 647 secondary antibody (green). Nuclei were counterstained with DAPI (blue). D Proximity ligation assay in NUP98-NSD1⁺ patient cells showing the interaction between the N-terminal NUP98 and C-terminal NSD1 moieties of the fusion with SMARCA5, detected using kit specific fluorophore (lem = 669 nm, far red), on the right. Nuclei were counterstained with DAPI (blue)