Nb110 59 906

5000 cells were seeded in laminin-coated 24-well plates. After fixation with 4% paraformaldehyde, cells were permeabilized with 0.1% Triton X100 for 15 min at room temperature. The cells were then incubated overnight at 4 ℃ with CIC antibody (NB110-59,906, NOVUS). Samples were rinsed three times in PBS and incubated for 1 h at room temperature with AlexaFluor 594 donkey anti-rabbit secondary antibody (Thermo Fisher Scientific). Nuclei were stained with 0.5 µg/ml DAPI for 1 min at room temperature. Images were acquired using fluorescence microscopy. The primary antibodies used for the immunoblots: anti-CIC (NB110-59,906, NOVUS), anti-ETV4 (10,684–1-AP, Proteintech), anti-xCT (NB300-318, NOVUS), anti-ACTB (A3854, Sigma), anti-GAPDH (2118, Cell Signaling).