LEF1 expression in B cells and B-cell derived lymphomas. (A) LEF1 protein expression in nuclear extracts of DLBCL cell lines was assayed by immunoblotting with LEF1 antibody using 293T and Raji cells as controls. Immunoblotting for Histone 3 served as a loading control. (B) LEF1 protein expression in cell extracts of normal B cells at different differentiation stages analyzed by immunoblotting with LEF1 antibody. (C) LEF1 mRNA expression in normal B cells at different differentiation stages and in primary CLL, FL, and DLBCL cell suspensions. (D) Immunohistochemistry of primary DLBCL specimens using LEF1 antibody. Representative examples of LEF1 negative (left) and positive (right) specimens are shown. Images were acquired using a Nikon Eclipse E1000 microscope (Nikon) equipped with 40× objective lenses. Images were captured with a SPOT flex mosaic 15.2 digital camera and software (Diagnostic Instruments). Digitized images were processed using Adobe Illustrator Version 10 software (Adobe Systems).