Figure 2
TDP-43 variants behavior in NSC34 and C2C12 cells. (a) Fractionation. NSC34 cells were transiently transfected with GFP-TDP-43, GFP-TDP-35, GFP-TDP-25. Western blot (WB) shows PBS, TRITON X-100, SDS and formic acid (FA) extracts, each set of sample loaded on a separate 10% polyacrylamide gel. Tubulin was used as loading control (full-length blots/gels are presented in Supplementary Figure S5). (b) Confocal microscope analysis. NSC34 cells were transiently transfected with GFP-TDP-43, or p2XFLAG-TDP-43. Nuclei were stained with DAPI. Images were acquired using confocal microscope with 63X magnification. (c) Nucleus/cytoplasm fractionation: NSC34 cells were transiently transfected with pcDNA3, GFP-TDP-43, or p2XFLAG-TDP-43. WB shows nuclear and cytoplasmic fractions, each set of sample loaded on a different 10% polyacrylamide gel (full-length blots/gels are presented in Supplementary Figure S5). (d) Filter retardation assay performed on PBS extracts of NSC34 transfected with pEGFP-TDP-43 and p2XFLAG-TDP-43 (full-length blots/gels are presented in Supplementary Figure S5). (e) Fractionation. C2C12 cells were transiently transfected with GFP-TDP-43, GFP-TDP-35, GFP-TDP-25. WB shows PBS, TRITON X-100, SDS and formic acid (FA) extracts, each set of sample loaded on a separate 10% polyacrylamide gel. Tubulin was used as loading control (full-length blots/gels are presented in Supplementary Figure S6). (f) Confocal microscope analysis. C2C12 cells were transiently transfected with GFP-TDP-43, or p2XFLAG-TDP-43. Nuclei were stained with DAPI. Images were acquired using confocal microscope with 63X magnification. (g) Nucleus/cytoplasm fractionation: C2C12 were transiently transfected with pcDNA3, GFP-TDP-43, or p2XFLAG-TDP-43. WB shows nuclear and cytoplasmic fractions, each set of sample loaded on a different 10% polyacrylamide gel (full-length blots/gels are presented in Supplementary Figure S6). (h) Filter retardation assay performed on PBS extracts of C2C12 transfected with pEGFP-TDP-43 and p2XFLAG-TDP-43 (full-length blots/gels are presented in Supplementary Figure S6).
