Figure 6. E-cadherin–dependent exosome... | Rockefeller University Press

Figure 6.

$Figure 6. E-cadherin–dependent exosome release of β-catenin. (A) CD82 interacts with the E-cadherin–β-catenin complex. HEK 293T cells stably expressing CD82 or empty vector were transfected with E-cadherin-GFP and β-catenin. 24 h after transfection, cells were lysed with the indicated lysis buffer and immunoprecipitated with anti-FLAG M2 affinity gel. The immunoprecipitated samples were washed with the indicated concentration of NaCl, followed by Western blotting with anti-GFP and anti–β-catenin antibodies (left). Total cell lysates were subjected to Western blotting with the indicated antibodies (right). (B) CD82 interacts with the E-cadherin–β-catenin complex in exosomes. Exosomes were purified from HEK 293T cells stably expressing CD82 or empty vector. Exosome fractions were lysed with 0.3% CHAPS lysis buffer and immunoprecipitated with anti-FLAG M2 affinity gel. The immunoprecipitated samples were washed with lysis buffer, followed by Western blotting with the indicated antibodies. (C) E-cadherin is required for CD82-induced exosome release of β-catenin. A431D cells were cotransfected with the indicated plasmids. After 48 h, total lysate and exosome fractions were purified and analyzed by Western blotting using the indicated antibodies.$

E-cadherin–dependent exosome release of β-catenin. (A) CD82 interacts with the E-cadherin–β-catenin complex. HEK 293T cells stably expressing CD82 or empty vector were transfected with E-cadherin-GFP and β-catenin. 24 h after transfection, cells were lysed with the indicated lysis buffer and immunoprecipitated with anti-FLAG M2 affinity gel. The immunoprecipitated samples were washed with the indicated concentration of NaCl, followed by Western blotting with anti-GFP and anti–β-catenin antibodies (left). Total cell lysates were subjected to Western blotting with the indicated antibodies (right). (B) CD82 interacts with the E-cadherin–β-catenin complex in exosomes. Exosomes were purified from HEK 293T cells stably expressing CD82 or empty vector. Exosome fractions were lysed with 0.3% CHAPS lysis buffer and immunoprecipitated with anti-FLAG M2 affinity gel. The immunoprecipitated samples were washed with lysis buffer, followed by Western blotting with the indicated antibodies. (C) E-cadherin is required for CD82-induced exosome release of β-catenin. A431D cells were cotransfected with the indicated plasmids. After 48 h, total lysate and exosome fractions were purified and analyzed by Western blotting using the indicated antibodies.

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