Rabbit anti matr3

Dissected Drosophila tissues or C2C12 cells grown on coverslips were rinsed in PBS (Lonza 17-512F) and fixed in 4% paraformaldehyde (Sigma P6148) for 20 min at room temperature. Following fixation, the samples were washed four times (×10 min) in PBS and blocked with blocking buffer: 5% normal goat serum (NGS; Abcam AB7681) in PBS with 0.1% TritonX-100 (PBST). The samples were incubated with primary antibody overnight at 4 °C, washed four times (x10 min) with 0.1% PBST, incubated with secondary antibody for 2 h at room temperature followed by 0.1% PBST washes. Samples were mounted onto slides using either ProLong^® Gold Antifade mounting reagent (Invitrogen P36930) or Fluoroshield (Sigma F6057).
Primary and secondary antibodies were prepared in blocking buffer.
Primary antibodies: Cy3-conjugated anti-HRP (1:100, Jackson ImmunoResearch 123-165-021); Rabbit anti-FLAG (1:500, Sigma F7425); Mouse anti-hnRNPM 2A6 (1:100; Santa Cruz sc-20001); Rabbit anti-MATR3 (1:500; Abcam ab151714)
Secondary antibodies: Goat anti-rabbit Alexa Fluor 488 (1:1000; Invitrogen A11008); Goat anti-mouse Alexa Fluor 546 (1:1000; Invitrogen A11030)

Motor neurons, matured for 50 days, and transiently transfected HEK293T cells at 72-h post transfection, were used for immunocytochemistry. Cells were washed and fixed with 4% paraformaldehyde and blocked for 1 h in Phosphate-buffered saline (PBS) containing 0.2% Triton X-100 and 10% normal donkey serum for iPSC-MNs and 5% normal goat serum for HEK293Ts respectively (Jackson ImmunoResearch). Samples were then incubated overnight at 4 °C with primary antibodies. The next day, cells were washed with PBS with 0.1% Triton X-100. Samples were then incubated with the appropriate secondary antibodies (diluted in blocking solution) conjugated to Alexa488, Alexa555 or Alexa647 fluorophores (1:500 to 1:1000 Molecular Probes) for 1 h at RT. Cell nuclei were labeled by DNA staining using DAPI or Hoechst (Life Technologies).
Primary antibodies Chicken anti-MAP2 (1:5000, Abcam ab5392); Goat anti-ChAT (1:500); Rabbit anti-MATR3 (1:200; Abcam 151714); Mouse Anti-digoxin (1:200; Jackson Immunoresearch 200-002-156); Rabbit anti-FLAG (1:500, Sigma F7425); Rabbit anti-Histone H3 (1:200; Abcam ab1791)

Nitrocellulose membranes were blocked in blocking buffer: 5% milk (BLOT- QuickBlocker^™ EMD Millipore WB57) in TBST followed by overnight incubation with primary antibody at 4 °C. Membranes were washed with TBST and incubated with secondary antibody for 1 h at room temperature, followed by washed with TBST. The membranes were imaged on Odyssey^® CLx (LI-COR Biosciences) and quantification of bands was performed using Image Studio™ (LI-COR Biosciences).
Primary and secondary antibodies were prepared in blocking buffer.
Primary antibodies: Mouse anti-FLAG (1:1000; Sigma F1804); Rabbit anti-MATR3 (1:4000; Abcam ab151714); Mouse anti-V5 (1:1000; Invitrogen R960-25); Mouse anti- α tubulin (1:8000; Sigma T5168)
Secondary antibodies: Goat anti-mouse Dylight 680 (1:10000; LI-COR 925-68070); Goat anti-rabbit Dylight 680 (1:10000; Invitrogen 35568); Goat anti-mouse Dylight 800 (1:10000; Invitrogen SA5-10176); Goat anti-rabbit Dylight 800 (1:10000; Invitrogen SA5-35571)