Effects of ITGB7 silencing on adhesion, migration, and invasion of MM cells. (A) Adhesion of calcein-AM–labeled ITGB7^silenced (ITGB7 shRNA2 and ITGB7 shRNA3) versus ITGB7^positive (scrambled shRNA) and parental (nontransfected) MM1S and H929 cells to BMSCs, FN, and E-CDH–coated 96-well microplates. Unattached cells were washed and adherent cells were measured in a fluorescence plate reader. Data are presented as percentage of respective controls (mean ± SD of triplicates from 3 independent experiments). (B) Transwell migration (8-μm pores; Costar) of calcein-AM–labeled ITGB7^silenced (ITGB7 shRNA2 and ITGB7 shRNA3) vs ITGB7^positive (scrambled shRNA) and parental (nontransfected) MM1S and H929 cells to RPMI serum-free media (cnt) or RPMI supplemented with SDF-1α (10 and 20nM). The fluorescence values, quantitated in a fluorescence multiwell plate reader using the 494/517nM filter set; percentage of migrating cells to SDF-1α versus control (serum-free RPMI) are shown. Data are presented as the mean ± SD of triplicates from 3 independent experiments. (C-D) Shown is a representative transwell Matrigel invasion of ITGB7^silenced (ITGB7 shRNA2 and ITGB7 shRNA3) vs ITGB7^positive (scrambled shRNA) and parental (nontransfected) MM1S and H929 cells under the conditions described in “Transwell migration assay and invasion studies.” Cells that invaded the Matrigel-coated filters are stained with crystal violet and counted using an inverted microscope. Images were acquired with a bright light Olympus BX5 microscope and multispectral camera (Nuance FX; CRi). 40× magnification.