Figure 2.

ADAP complex and integrin activation. (A) ADAP is involved in the dendritic expression of activated β1-integrin in hippocampal neurons (DIV9). Antibody 9EG7 detects activated β1-integrins on the soma and neurites; reduced labeling intensity is evident in cells with shRNA-mediated knock down of ADAP and recovery to control levels is seen upon re-expression of a knock-down resistant ADAP clone. Scale bar, 100 μm (B) Quantitative analysis of labeling intensity along dendrites confirms the reduction of 9EG7 labeling upon ADAP knock down as well as its rescue (n = 24–30 per condition). (C) Effective knock down of murine ADAP and its rescue through re-expression of the shRNA-resistant human form (hADAP) in HEK-293T cells. (D,E) In contrast to the activated form, total β1-integrin was increased on the dendrites following ADAP knock down, indicating that the observed reduction in dendritic labeling with antibody 9EG7 is related to reduced β1-integrin activation by conformational change rather than changes in expression level. Data are Mean ±SEM. *p < 0.01.