Fig. 2

Numbl interacts with Integrin β1. a The interaction between endogenous Numbl and Integrin β1 in myeloma cell lysate was assessed by immunoprecipitation with an anti-Integrin β1 antibody or with a mouse normal IgG and analyzed by Western blot analysis using anti-Numbl antibody. b HA-tagged Numbl and GFP-tagged Integrin β1 were co-expressed in HEK293T cells. Extracts with equal amount of proteins were immunoprecipitated with anti-HA or anti-GFP antibodies and analyzed by immunoblotting with anti-GFP or anti-HA antibodies. c Co-localization of Integrin β1 and Numbl. The HA-Numbl and GFP-Integrin β1 plasmids were co-transfected into RPMI 8226 and HEK293T cells. After 48 h, both cells were visualized by confocal fluorescent microscopy using Hoechest 33,342 for nucleus staining. The right panel (Merge) shows the merging of all three panels (images taken with X40 magnification). d The quantification of images from C. A minimum of 200 cells per sample were counted, and the percentage of cells with Numbl and Integrin β1-double positive cells was calculated. Results represent the means of data from 3 independent experiments