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i.p : GFP- Tspan blot: GFP- Tspan

Tspan14. Tspan12. CD9. -. GFP- Tspan :. +. +. +. +. HA-ADAM10:. A. i.p : GFP- Tspan blot: HA-ADAM10. **. 125. 80. 80. 100. HA-ADAM10 quantitation from GFP i.p . Relative ADAM10 level. 75. 58. 58. 58. 50. 25. 0. B. whole cell lysate blot: HA-ADAM10. **. 60. 40.

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i.p : GFP- Tspan blot: GFP- Tspan

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  1. Tspan14 Tspan12 CD9 - GFP-Tspan: + + + + HA-ADAM10: A i.p: GFP-Tspan blot: HA-ADAM10 ** 125 80 80 100 HA-ADAM10 quantitation from GFP i.p. Relative ADAM10 level 75 58 58 58 50 25 0 B whole cell lysate blot: HA-ADAM10 ** 60 40 HA-ADAM10 quantitation from whole cell lysate % mature ADAM10 20 0 46 C 30 i.p: GFP-Tspan blot: GFP-Tspan 25 Supplementary Figure III.The non-TspanC8 tetraspanin Tspan12 does not interact with or promote ADAM10 maturation. The experiment was performed and presented as described in Figure 2A-C, with the exception that GFP-tagged mouse tetraspanins were used instead of FLAG-tagged tetraspanins. The GFP-tagged mouse forms of non-TspanC8 tetraspanins CD9 and Tspan12 were compared to Tspan14 in their capacity to (A) co-immunoprecipitate with, and (B) promote maturation of, bovine HA-tagged ADAM10 (n=5; **p<0.01 compared to all other data; error bars represent standard errors of the mean). The GFP immunoprecipitation was performed with GFP-Trap beads and confirmation of similar Tspan12 and Tspan14 expression was obtained by (C) anti-GFP western blotting.

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