Fig. 5. Immunohistochemistry for ATBF1 (red, A,B) counterstained with the general membrane dye DiOC₆(3) (green, A,B) in RA-treated P19 cells. (A) Subcellular localization of ATBF1 in the adhesive condition. Nuclear localization of ATBF1 on dishes coated with poly-L-onetime (A1), poly-L-ornithine+fibronectin (A2), poly-L-ornithine+laminin (A3), poly-L-lysine (A4) and gelatin (A5) at 3 hours after cell transfer to the dishes. Cells pre-treated with LY294002 (20 µM, A6), ryanodine (100 nM, A7) and caffeine (5 mM, A8). (B) Subcellular localization of ATBF1 in floating conditions. ATBF1 is localized in the cytoplasm of cells in the aggregated and floating conditions (B1). Treatment with leptomycin B (10 nM, 1 hour, B2) induces ATBF1 accumulation in the nucleus in the floating condition. ATBF1 is localized in the nucleus of cells separated from embryonic bodies after treatment with EDTA (1 mM, 30 minutes) and pipette (B3).