Figure 3.

TDP-43 is cleaved at Q327. (a) HeLa Cells transiently transfected with HA-TDP-43-GFP (WT) or HA-TDP-43^Q327L-GFP (Q327L) were either sham or CVB3-infected for 7 h. Cells were collected for western blot analysis of the expression of TDP-43 using anti-HA antibody, VP1, and β-actin. Densitometric analysis was carried out as in Figure 1. (b) Fifty microgram of protein extracts from HeLa cells transiently expressing WT or HA-TDP-43^Q327L-GFP was incubated with increasing concentrations of viral protease 3C or 2A as indicated for 8 h. CVB3-infected HeLa lysates were used as a control to compare the generation of similar 35 kDa cleavage fragments. Protein expression of TDP-43, VP1, and β-actin was detected as described above. * indicates a cleaved TDP-43 band at ~35 kDa. (c) Schematic diagram of the functional domains, the identified cleavage site, and the resulting cleavage fragments of TDP-43. RRM, RNA-recognition motif; NLS, nuclear localization signal; and NES, nuclear export signal