Fig. 1

P-12-lipoxygenase interacts with integrin β4 subunit in vitro. (a) A431 cells were stimulated with mAb β4 3E1 and harvested at timed intervals from 5–90 min. 12-LOX and the β4 subunit co-immunoprecipitated from untransfected A431 cells (A431), [and A431 cells over expressing 12-LOX (Tx-A431)-Additional file 1]. (b) CHO cells were transfected with different β4 constructs either alone or in combination with 12-LOX: (1) control: vector alone; (2) CHO: nontransfected cells; (3) Txβ4: full-length β4; (4) Txβ4 + 12-LOX: cotransfected with full-length β4 and 12-LOX; (5) Txtruncated1: pCMV-β4 Δ70-660 (c-myc tagged head-less) +12-LOX; (6) Txtruncated2: pCMV-β4 Δ854-1752 (tail-less) +12-LOX. 12-LOX and the β4 subunit coimmunoprecipitated from three transfectants: Txβ4, Txβ4 + 12-LOX, and Txtruncated1 (headless). Positions of 12-LOX and β4 are indicated. The experiment was repeated three times. For each experiment, mouse or rabbit IgG (control 1) and Sepharose 4B-conjugated protein G beads alone (control 2) were used as negative controls. (C) 12-LOX and the β4 subunit co-immunoprecipitated from A431 cells after growth on laminin at the same timed interval as with mAb 3E1 stimulation. Whole cell lysate of A431 stimulated with mAB 3E1 was loaded as a control (A431-3E1T). β4 subunit was detected with mAb 450-11A