Fig. 3: Electron microscopy analysis reveals abnormal NM morphology in N2a cells containing TDP-CTF aggregates.
From: TDP-43 pathology disrupts nuclear pore complexes and nucleocytoplasmic transport in ALS/FTD
a, Schematic domain structures of engineered peroxidase, APEX2, fusion TDP-43 and TDP-CTF. b, Nuclear signals of Flag-APEX2-TDP-43 and cytoplasmic aggregates of Flag-APEX2-TDP-CTF were detected by anti-Flag antibody (green). TDP-CTF phosphorylation was detected by anti-pTDP-43 (Ser409/Ser410) antibody (red). Protein biotinylation catalyzed by APEX2 was detected by Cy5-conjugated streptavidin (magenta). Scale bar: 10 μm. c, Electron microscopy of untransfected N2a cells and cells expressing Flag-APEX2-TDP-43, Flag-APEX2-TDP-CTF or GFP-TDP-CTF. Untransfected cells showed dense chromatin and round nuclei. APEX2 catalyzed the deposition of diaminobenzidine (DAB) near nuclear APEX2-TDP-43 and in cytoplasmic APEX2-TDP-CTF aggregates. APEX2-TDP-CTF- as well as GFP-TDP-CTF-expressing cells exhibited irregularly shaped nuclei with deep invaginations of the NM. Each experiment was repeated independently three times with similar results.
