Re blot plus buffer

Cells were lysed in radioimmunoprecipitation assay (RIPA) buffer (Fisher Thermo Scientific) supplemented with complete Mini EDTA-free protease inhibitor cocktail (Roche), at 1 × 10⁶ cells in 100 μl of lysis buffer. Postnuclear lysates were resolved by sodium dodecylsulfate–polyacrylamide gel electrophoresis using 4–15% BisTris NuPAGE gels (Invitrogen) and proteins were transferred to membranes (Immunoblot PVDF membranes, BioRad). Membranes were stained with primary antibodies against ETV6/Tel (Novus Biologicals, catalog no. NBP1-80695, 0.4 μg ml⁻¹), ETV3 (Atlas Antibodies, catalog no. HPA004794, 0.4 μg ml⁻¹), GP96 (Novus Biologicals, clone 9G10, 0.4 μg ml⁻¹) or actin (Millipore, clone C4, 0.4 μg ml⁻¹), followed by horeseradish peroxidase-conjugated secondary antibodies (Jackson Immunoresearch, dilution 1:10,000). Some membranes were incubated with Re-blot Plus buffer (Millipore). Densitometry quantification was performed using Fiji (v.2.9).

Cells were lysed in RIPA buffer (Thermo Scientific) supplemented with complete Mini EDTA‐free protease inhibitor cocktail (Roche), at 1 × 10⁶ cells in 100 μl of lysis buffer. Post‐nuclear lysates were resolved by SDS–PAGE using 4–15% BisTris NuPAGE gels (Invitrogen) and proteins were transferred to membranes (Immunoblot PVDF membranes, Bio‐Rad). Membranes were stained with primary antibodies against IRF1 (Cell Signaling, clone D5E4), ZNF366 (Novus Biologicals, polyclonal, reference AF4707), MAFF (Novus Biologicals, polyclonal, reference AF3917), or actin (Millipore, clone C4), followed by HRP‐conjugated secondary antibodies (Jackson Immunoresearch). Some membranes were incubated with “Re‐blot Plus” buffer (Millipore).