FIGURE 11.

Differential effects of TspanC8s on ADAM10 substrate cleavage: Tspan15 promotes cleavage of N-cadherin and Tspan14 reduces cleavage of GPVI. A, HEK-293T cells were mock transfected (−) or transfected with FLAG-tagged mouse TspanC8s. The cells were lysed in 1% Triton X-100 lysis buffer and subjected to Western blotting with an antibody to the C-terminal cytoplasmic tail of N-cadherin (upper panel) or with an antibody to the FLAG epitope (lower panel). B, data from A (upper panel) were quantitated and the lower, cleaved band given as a percentage of the total (upper and lower band combined). Data were normalized by log transformation and statistically analyzed using a one-way ANOVA with a Dunnett's multiple comparison test compared with the mock control. Error bars represent the standard error of the mean from three experiments (*, p < 0.05). C, HEK-293T cells were co-transfected with GPVI and FcRγ and one of each of the FLAG-tagged mouse TspanC8s or without a tetraspanin (−) or with the addition of the ADAM10 inhibitor GI254023X at 10 μm. Cells were treated as in panel A, except lysates were subjected to an anti-GFP antibody (upper panel) instead of an anti-N-cadherin antibody. D, data from panel C (upper panel) were quantitated as described in panel A (***, p < 0.001).