Figure 3. Complete loss of neurofilament light (NEFL) protein in cultured patient neurons.

(A) Immunoblotting of whole cell lysates of patient 1 clones 1 and 2 (Pt C1 and C2) and controls 1-3 (ctr 1-3) after motor neuronal differentiation with an N-terminal monoclonal or a polyclonal pan-NEFL antibody. Protein levels of neuronal markers ChAT, TUJ1, and neurofilament medium (NEFM) and the loading control glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as well as the stain-free blot are shown. (B) Immunocytochemical analysis of NEFM (green) and NEFL (orange) of neurite architecture in patient 1 and control neurons after motor neural differentiation. 4′,6-diamidino-2-phenylindole (DAPI) indicates nuclear staining. Scale bars 50 μm.