Figure 1

TDP-43 and FUS/TLS aggregate at nuclear NEAT1_2 foci. A. At 48 hours after transfection with wild-type (WT) and 35-kDa TDP-43 fragment and WT FUS/TLS with the V5 tag at the C terminus, HeLa cells were fixed with 4% paraformaldehyde, hybridized with fluorescein isothiocyanate (FITC)-labeled RNA probe against NEAT1_2 long non-coding RNA (lncRNA), and double-immunostained with polyclonal anti-FITC and monoclonal anti-V5 antibodies. Schematic diagram (right) represents the TDP-43 isoforms and FUS/TLS. NLS, Nuclear localization signal; RRM, RNA recognition motif; C-term, C-terminal domain; GR, glycine-rich motif; NES, nuclear export signal; RGG, arginine-glycine-glycine motif ; ZF, zinc-finger motif. Dotted lines represent the outline of the nucleus. Scale bars, 10μm. B. The Y-axis shows what percent NEAT1_2 foci is overlapped by TDP-43 nuclear aggregates in (A). The 26-kDa TDP-43 fragment hardly accumulates in nuclei and lacks affinity for NEAT1_2 foci. Data represent mean ± s.d. (n = 50 for each transfection of TDP-43 isoforms). *P < 0.0001 (unpaired Student’s t-test).