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. 2013 Aug 23;14(10):884–890. doi: 10.1038/embor.2013.128

Figure 5.

Figure 5

iRhoms are specific to TACE and are essential for stimulated TACE shedding. (A) Western blot of endogenous ADAM9, ADAM10 and TACE from ConA-enriched lysates from WT or iRhom DKO MEFs. Immature and mature ADAMs are indicated by black and white arrowheads, respectively. (B) Western blot showing tagged ADAMs −7, −8, −12, −15, −19, −22, −23 and −33 expressed in WT and iRhom DKO MEFs. Immature and mature ADAMs are indicated by black and white arrowheads, respectively. The immature and mature forms of ADAM7 can only be revealed by deglycosylation (see supplementary Fig S2A online). The panel for ADAM23 shows three bands: the upper band (asterisk) is the Golgi species with prodomain intact; the middle band (black arrowhead) is the immature ER form; the lower band is mature ADAM23 lacking the prodomain (white arrowhead). (C) Shedding of luciferase-HB-EGF after stimulation with 250 nM phorbol myristate acetate for 1 h. The mean −/+ s.d. of three independent experiments is shown.*P<=0.01, t-test. (D) Shedding of luciferase-HB-EGF on stimulation with 1 μM bombesin for 1 h. The mean −/+ s.d. of three independent experiments is shown.**P<0.05, t-test. (E) Shedding of unmodified secreted luciferase by WT versus iRhom DKO MEFs. The mean −/+ s.d. of three independent experiments is shown. DKO, double KO; KO, knockout; MEFs, mouse embryonic fibroblasts; WT, wild type.