Fig. 4. Alteration of the subcellular localization of SIAH1 in cotransfection with BOB.1/OBF.1. NIH 3T3 cells were transfected with a BOB.1/OBF.1 construct containing an additional NLS sequence (A), Flag-SIAH (B) or both together (C and D). 18 h after transfection, the cells were fixed and stained with polyclonal anti-BOB.1/OBF.1 antibody and FITC-conjugated goat anti-rabbit antibody (A), monoclonal anti-FLAG antibody M2 and Cy3-conjugated goat anti-mouse antibody (B), or with both first and secondary antibodies together (C and D).