Functional recombinant N-cadherin domains 1 and 2 block KLRG1 binding to E-cadherin. (A) SDS-PAGE of recombinant N-cadherin. Extracellular domains 1 and 2 of N-cadherin were produced and purified by Ni²⁺ affinity chromatography. Lane 1: lysate; lane 2: purified N-cadherin. The protein bands were revealed using Gel Code Blue staining reagent (Coomassie). (B) KLRG1 tetramer and (C) anti-TCRβ mAb were first incubated 30 minutes with N-cadherin before incubation with DO11 E-cadherin WT cells. Cells were washed 3 times and analyzed by FACS. The results are representative of 5 independent experiments. (D) N-cadherin– or BSA-coated plates were incubated with parental or KLRG1 reporter cells. Fixed cells were washed and treated with X-Gal substrate and imaged at ×10 with an Olympus DP70. The results are representative of 3 independent experiments.