HSPH1 interaction with c-Myc and Bcl-6 in human aggressive B-NHLs. (A) Western blot analyses of HSPH1 and Bcl-6 (left) or HSPH1 and c-Myc (right) in Namalwa whole-protein extract immunoprecipitated (IP) with Abs against the indicated molecules or control isotype IgG. Whole lysate was included as an internal control. For c-Myc immunoprecipitation, proteins were eluted in nonreducing condition to avoid the detection of Ab heavy chains in the region in which c-Myc protein migrates. Western blot images were acquired using Microtek ArtixScan F1 and cropped to retain the relevant bands with Adobe Photoshop CS version 4 for Macintosh computer. (B) Representative microphotographs of double-marker immunofluorescence for HSPH1 (green signal) and either Bcl-6 or c-Myc (red signal) in 2 cases of germinal center–type DLBCLs with (middle) or without (left) c-Myc amplification, and 1 case of BL (right). Bindings of primary rabbit anti-human HSPH1 Ab and mouse anti-human Bcl-6 (clone PG-B6p) or anti-human c-Myc (clone 9E10) were revealed, respectively, by an Alexa-488– and an Alexa-568–conjugated secondary Abs. Sections were analyzed under a Leica DMRBE microscope and microphotographs were collected with a Leica MC120HD digital camera.