Mouse anti yh2a x

The following antibodies were used: mouse anti-yH2AX (Millipore), mouse anti beta-actin (Abcam) and IRDye 680RD donkey anti-mouse (LI-COR), rabbit anti-cleaved caspase 3 (Abcam), mouse anti y-H2AX (Millipore), anti-rabbit/555 (Invitrogen) and anti-mouse/488 (Life technologies). Chemicals were purchased from Sigma-Aldrich. MTH1 inhibitors were synthesized in-house and dissolved in DMSO to 10 mM [10 (link), 12 (link)].

For immunofluorescence staining, cells were washed twice with PBS without Ca²⁺/Mg²⁺ (LifeTechnologies) and fixed with 4% PFA in PBS for 10 min at room temperature. PFA was aspirated and cells were washed three times with PBS. Fixed cells were first permeabilized for 10 min in 0.2% Triton X solution and subsequently incubated for 1 h at RT in blocking solution (1% BSA, 5% donkey serum, 0.3 M glycine and 0.02% Triton X in PBS). Following blocking, primary antibodies were diluted in blocking solution and cells were incubated with primary antibody solution overnight at 4 °C except for the γH2A.X antibody which was kept for only 2 h at room temperature on the fixed material. The following primary antibodies were used: chicken anti-SMI32 (1:10,000, Covance), mouse anti-FUS (1:5000, Sigma-Aldrich), rat anti-meFUS (1:100^{5 (link)},), rabbit anti-beta-III-Tubulin (1:3000, Covance), mouse anti-Hb9 (1:100 Development studies Hybridoma Bank), rabbit anti-Islet (1:500, Abcam), mouse anti-yH2A.X (1:500 Millipore), rabbit anti-ChAt (1:500, Chemicon), rabbit anti-53BP1 (1:1000, Novusbio). Nuclei were counter stained using Hoechst (LifeTechnologies).