46

2530

i.p: GFP-Tspan

blot: GFP-Tspan

C

% m

atur

e A

DA

M10 60

40

20

0

HA-ADAM10 quantitation

from whole cell lysate

58

**

whole cell lysate

blot: HA-ADAM108058

B

Rel

ativ

eA

DA

M10

leve

l

HA-ADAM10 quantitation

from GFP i.p.

1007550250

125

i.p: GFP-Tspan

blot: HA-ADAM108058

A

Tspa

n14

-

+ + + +GFP-Tspan:

HA-ADAM10:

CD

9

Tspa

n12

**

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.