Differential protein expression of BIN1 and MYC in AHI-1–suppressed and –overexpressed CTCL cells. (A) Up-regulation of BIN1 was evident in AHI-1–suppressed Hut 78/sh4 cell lines. Of the 3 isoforms detected at 65, 60, and 55 kDa, the most significant up-regulation was observed in the 65- and 60-kDa isoforms. Reduced BIN1 protein expression was also observed when AHI-1 was overexpressed in Hut 78 cells (Hut 78 AHI-1). (B) Quantification of the differential expression of the 65-kDa isoform of BIN1 relative to actin. (C) Changes in MYC expression were observed in AHI-1–suppressed Hut 78/sh4 cell lines and AHI-1–overexpressed cells compared with the control cells (Hut 78 and Hut 78 RPG). Vertical line has been inserted to indicate a repositioned gel lane. (D) Quantification of the differential expression of MYC relative to actin. (E,F) Protein lysates from control Hut 78 cells, AHI-1–suppressed cells (sh4 bulk), and AHI-1–overexpressed cells were used to immunoprecipitate BIN1 (E) and MYC (F), and proteins were detected by Western blotting with antibodies to MYC (E) and BIN1 (F) as indicated. Values shown are the mean plus or minus SEM. The P values presented are the result of 2-sample t tests comparing the AHI-1–suppressed expression data with that of Hut 78 controls.