Sc 1751

For doing western blots in Drosophila, the following primary antibodies were used: anti-FUS (Bethyl Laboratories; A300-302A, 1:2000), anti-MBNL1 (Millipore; MabE70, 1:2000), and anti-mbnl1 (Invitrogen; MA1-16967, 1:1000). DyLight fluorescent secondary antibodies (Thermo Fisher Scientific) were used at a concentration of 1:10,000
For the Drosophila NMJ analysis, the following primary antibodies were used: Alexa Fluor 488-conjugated goat anti-HRP (Jackson Immuno Research; 123-545-021, 1:200) and mouse anti-DLG 4F3 (DSHB, 1:100). The following secondary antibodies were used: Alexa Fluor 647-conjugated anti-Phalloidin (Invitrogen; A22287, 1:250) and goat antimouse Alexa Fluor 546 (Invitrogen, A11030, 1:500).
The following primary antibodies were used: anti-G3BP1 (Protein Tech; 13057-2-AP, 1:2000) and anti-HA7 (Sigma, H3663, 1:500). Alexa-Fluor fluorescent secondary antibodies (Invitrogen) were used at a concentration of 1:500 for doing Western blots in HEK293 and N2a cell lines.
The following primary antibodies were used for doing immunoblotting in primary neuronal cells: anti-MAP2 (EMD Millipore; AB5622, 1:500), anti-FUS (Proteintech; 11570-1-AP, 1:200), anti-TIA1 (Santa Cruz; SC-1751, 1:250), anti-G3BP1 (Proteintech; D5444, 1:100), anti-MBNL1 (Millipore; MABE70, 1:100), and anti-NEFL (Novus Biologicals; NB300-222, 1:300).

Tissue sections were deparaffinized in xylene and rehydrated through a series of ethanol solutions. Antigen retrieval was performed by steaming in sodium citrate buffer (10 mM sodium citrate, 0.05% Tween-20, pH 6) for 30 min. Tissues were immunostained with antibodies against poly(GR) (1:1000, EMD Millipore, MABN778), TIA1 (1:50, Santa Cruz Biotechnology, sc-1751), ataxin-2 (1:500, Proteintech, 21,776–1-AP), and/or pTDP-43 (1:1000, Cosmo, CAC-TIP-PTD-P02) overnight at 4 °C. Sections were subsequently incubated with corresponding secondary antibodies (1:500, Molecular Probes) and coverslipped with Vectashield with DAPI. Images were taken with Zeiss LSM 800 confocal microscope.