A) Cartoon of JPH2 showing location of the NLS, the G482 cleavage site, and the N-terminal MYC and C-terminal HA tags. B) Confocal images of HEK293 cells expressing dual-tagged JPH2 stained with MYC (red), HA (green) antibodies, and DAPI (blue). Green staining shows that JPH2-CTP translocated into the nucleus. C) Alignment of a putative NLS in JPH2 across species. D) Confocal images of HEK293 cells expressing dual-tagged JPH2-NLS-5A (mutated NLS peptide) stained with MYC (N-terminus, red) HA (C-terminus, green), and DAPI (nucleus, blue) antibodies. Lack of green staining shows that the NLS is required for nuclear translocation of JPH2-CTP. Dotted white box indicates magnified region (3x zoom); scale bar is 10μm. All the experiments were performed at least 3 times to validate reproducibility.