Figure 1.

Expression of FBXO25 protein in mouse tissues and cultured cell lines. Approximately 150 μg of protein from the indicated tissues (A) and cultured cells (B) lysates were subjected to SDS-PAGE, transferred onto nitrocellulose membranes, and probed with affinity-purified anti-FBXO25 antibodies (1:1500). Tissue and cell extracts were prepared as described in Materials and Methods. The specificity of the anti-FBXO25 antibodies (C) were ascertained by probing twin blots prepared from SDS-PAGE loaded with two separate sets of cell lysates from HEK293H cells and HEK293H cells stably transfected with GST-FBXO25 (HEK293H^FB25-WT-1), with antibodies in the absence and presence of 6 μg/ml recombinant FBXO25 N-terminal fragment. Note that the FBXO25 fragment fully blocked the antibody reaction with both the endogenous and GST-fusioned protein. Ponceau-S staining showed protein loading (middle) and anti-GST antibodies labeling showed the expression of GST-FBXO25 protein in HEK293H^FB25-WT-1 cells (bottom).