ITGB7 silencing reduces p65 NF-κB activity in MM cells. (A) NF-κB-p65 transcription factor binding to its consensus sequence on a plate-bound oligonucleotide was measured by ELISA in nuclear extracts from Jurkat, ITGB7^silenced (ITGB7 shRNA2 and ITGB7 shRNA3), and ITGB7^positive (scrambled shRNA) MM1S and H929 cells cultured on FN-coated plates. Shown results represent means (± SD) of triplicate experiments (* P < .05). (B) Decreased nuclear NF-κB-p65 translocation or localization in ITGB7^silenced (right) compared with ITGB7^positive (left) MM1S cells cultured on FN-coated plates as detected by immunofluorescence staining with p65 NF-kB Ab (Cy-3 labeled) and DAPI for nuclear visualization. Image acquisition was performed with epifluorescence Olympus BX5 microscope and multispectral color camera (Nuance FX; CRi) with 60× magnification and oil immersion (details in supplemental Materials).