Figure 3.

iRhom2 Cytoplasmic Tail Phosphorylation Is Required for Rapid Induction of TACE-Dependent Shedding

(A) iRhom2 Dead mutant stably expressed in HEK293ET cells is not phosphorylated in response to PMA.

(B) PMA-stimulated TGF-α-AP shedding requires iRhom2 phosphorylation. iRhom1/2 DKO MEFs were transduced with iRhom2 WT, iRhom2 Dead mutant, or empty vector (EV) retrovirus.

(C) Marimastat (MM; 5 μM) was used to inhibit shedding by metalloproteases.

(D) The ADAM10-specific inhibitor GI254023X (GI; 1 μM, 60 min) did not affect TGF-α-AP shedding in rescue assays in iRhom DKO MEFs expressing WT iRhom2 versus the Dead mutant or empty vector.

(E) Ionomycin (IO; 2.5 μM, 60 min) stimulated shedding of betacellulin-AP (BTC-AP) was unaffected by iRhom2 phosphorylation.

(F) iRhom2 KO BMDMs transduced with iRhom2 Dead retrovirus are defective in the shedding of TNF after 3 hr of LPS stimulation.

Data are presented as mean ± SEM. See also Table S1.