Fig. 10. Altered extractability of ß1 integrin in Triton X-100 in 3T3-L1 cells overexpressing ADAM12 at the cell surface. Adherent 3T3-L1 ADAM12-cyt cells and control 3T3-L1 cells treated with 0.01% Triton X-100 in DMEM (A,B) for 5 minutes on ice or left untreated (C,D) and subsequently immunostained with polyclonal antibodies to ß1 integrin (A-D). In parallel experiments, adherent 3T3-L1 ADAM12-cyt cells (E) or control cells (F), or normal growing 3T3-L1 cells (G) or confluent day 0 3T3-L1 cells (H) were extracted with 0.5% Triton X-100 in a physiological buffer (CSK) for 5 minutes on ice, released from the plastic substrate with dissociation buffer and immunostained with polyclonal antibodies to ß1 integrin in suspension as described in Materials and Methods. Note that the increase in ß1 integrin solubility, and hence, decreased immunostaining in cells overexpressing ADAM12-cyt (A,E) is similar to that seen in normal confluent day 0 3T3-L1 cells (H) compared with normal growing 3T3-L1 cells (G). Bars, 25 µm (A-D) and 15 µm (E-H).