Fig. 5

TDP-43 interacts with PSF and NeuN. a Silver-staining of 4-12 % SDS-PAGE gel showing GST-TDP-43 pull-down from pig cortex lysates. GST served as a control. The indicated band was analyzed by mass spectrometry and found to contain PSF. b Western blotting with anti-PSF verifying the interaction of GST-TDP-43 and PSF in pig brain cortex lysates. Two GST and GST-TDP-43 pull-down proteins were analyzed. c Purified His-PSF was incubated with transgenic pig brain cortex lysates, and the bound proteins were subjected to western blot, which shows the in vitro direct interaction of endogenous pig TDP-43 and transgenic human TDP-43 with His-PSF. Input is 5 % of purified His-PSF, and 40 % of pulldown was loaded into the SDS gel. d PSF contains N-terminal, RRM1, RRM2, and C-terminal domains. The N-terminal domain is a glycine/proline/arginine rich domain, the C-terminal and the NOPS regions of PSF were found to bind to TDP-43. TDP-43 contains N-terminal, RRM1, RRM2, and C-terminal (G-rich) domains. e Purification of His-PSF containing different domains as indicated with the corresponding amino acid positions. f In vitro GST-pull down identifying that the C-terminal regions containing amino acids 444–707 of PSF bind to C-terminal GST-TDP-43 (amino acids 264–414), but not N-terminal TDP-43 (amino acids 1–251)