Fig. 2. A clathrin-binding region within Dab2. Approximately 50 µg of either GST (lanes a and b) or GST–Dab2(206–258) (lanes c and d), GST–Dab2(206–258) (LVD→AAA) (lanes e and f), GST–Dab2(206–368) (lanes g and h), GST–Dab2(206–368) (LVD→AAA) (lanes i and j) or GST–Dab2(206–368) (LVD→AAA/W→A) (lanes k and l) immobilized on GSH–Sepharose were incubated with rat brain cytosol. After centrifugation, aliquots corresponding to 1/50 of each supernatant (S) and 1/5 of each washed pellet (P) were resolved by SDS–PAGE and either stained with Coomassie Blue or transferred to nitrocellulose. Portions of the blots were probed with the anti-AP-2 α-subunit mAb 100/2, anti-AP-2 µ2-subunit antiserum, the anti-clathrin HC mAb TD.1 or the anti-clathrin LC mAb Cl 57.3.