Immunofluorescence Assay for TDP-43 Localization

For these assays, 293 T cells were transfected with or without plasmids and then fixed with 3.7% paraformaldehyde in PBS at RT for 15 min at 24 h (for GFP-TDP-43-IIPLD experiments) or 48 h post-transfection. The fixed cells were incubated with a TDP-43 or lamin A/C or His antibody at 4 °C overnight, followed by incubation with a secondary antibody conjugated with a fluorescent dye (Molecular Probes). The staining cells were gently fixed with 3.7% paraformaldehyde for 2 min before mounting. Slides were mounted using Vectashield DAPI H-1200 (Vector Laboratories). Cellular fluorescence images were obtained from a single optical section using an LSM710 META laser confocal microscope (Zeiss) or ELYRA super-resolution microscope (Zeiss)^{33 (link)}.