FIGURE 8.

Cleavage of TDP-43 leads to attenuation of TDP-43-induced death. A, NSC34 cells, seeded on 6-well plates at 1 × 10⁵ cells/well, were co-infected with the indicated virus at a m.o.i. of 400 in association with cre-recombinase or LacZ virus at a m.o.i. of 40. 24 h after infection, cell medium was replaced with DMEM/N2 supplement with or without 2 μm MG132. 24 h after medium replacement, LDH release from cells was measured. B, immunoblot (IB) analyses were performed with antibodies to cleaved caspase-3, Bim, Bcl-xL, and TDP43-C. #, GAPDH band. C and D, U2OS cells, seeded on 6-well plates at 5 × 10⁴/well, were co-infected with TDP-43-(1–414) or TDP-43-(170–414) virus at a m.o.i. of 100 for TDP-43-(1–414) or 800 for TDP-43-(170–414) in association with cre-recombinase or LacZ virus at a m.o.i. of 40. 24 h after infection, cell medium was replaced with DMEM/N2 supplement. 24 h after medium replacement, LDH release was measured (D). Immunoblot analysis was performed with antibodies to TDP43-C (C). <, TDP-43-(1–414); ≪, TDP-43-(170–414). E and F, NSC34 cells, seeded on 6-well plates at 1 × 10⁵/well, were co-infected with TDP-43-wt or -D89E/D169E virus at a m.o.i. of 400 in association with cre-recombinase or LacZ virus at a m.o.i. of 40. 24 h after infection, cell medium was replaced with DMEM/N2 supplement. 24 h after medium replacement, LDH release was measured (F). Immunoblot analysis was performed with antibody to TDP43-C (E). <, CTF35; #, an uncharacterized protein, migrating slightly slower than CTF35. exp, exposure. G and H, NSC34 cells, seeded on 6-well plates at 1 × 10⁵ cells/well, were co-infected with an indicated adenovirus at a m.o.i. of 400. All samples were co-infected with cre-recombinase virus at a m.o.i. of 40. 24 h after infection, cell medium was replaced with DMEM/N2 supplement. 24 h after medium replacement, LDH release from cells was measured (G). Immunoblot analysis was performed with antibodies to cleaved caspase-3, Bim, Bcl-xL, and TDP43-C (H).