Skip to main content
. 2023 Feb 9;299(3):102995. doi: 10.1016/j.jbc.2023.102995

Figure 4.

Figure 4

FUS and TDP-43 modulate poly(GR) expression through the mTORC2/AKT/VCP axis.A and B, Western blot analysis showing effect of various genetic manipulations of RQC factors (A) or mTORC2/AKT pathway components (B) on the suppression of Flag-(GR)80 expression by FUS-RFP in Mhc>Flag-GR80/FUS-RFP flies. C and D, Western blot analysis showing effect of knockdown of AKT (C) or VCP (D) on the suppression of Flag-(GR)80 expression by TDP43-RFP in Mhc>Flag-GR80/TDP43-RFP flies. E, effect of FUS and TDP-43 on AKT phosphorylation level in Mhc>Flag-(GR)80 flies. F, dot blots showing effect of FUS and TDP-43 on poly(GR) expression in C9ALS patient fibroblasts. G, effect of combined RNAi of AKT1, AKT2, and AKT3 on Flag-GR80 expression in HEK293T cells with or without Flag-FUS or Flag-TDP43 coexpression. Plasmid expressing GFP is used a transfection control. Bar graphs show quantification of the relative expression of (GR)80 in all panels (∗∗∗p < 0.001; ∗∗p < 0.01). ALS, amyotrophic lateral sclerosis; FUS, fused in sarcoma; HEK, human embryonic kidney; mTORC2, mechanistic target of rapamycin complex 2; RQC, ribosome-associated quality control; TDP-43, TAR DNA-binding protein 43; VCP, valosin-containing protein.