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. 2013 Jun 25;110(28):11433–11438. doi: 10.1073/pnas.1302553110

Fig. 5.

Fig. 5.

Effect of inactivation of iRhom1 or iRhom2 or both iRhom1 and -2 on ADAM17- and ADAM10-dependent shedding. iRhom2−/− mEFs (A, E, G, H, I, and K) or WT controls (B, F, and J) were transfected with the AP-tagged ADAM17 substrates TGFα (A, B, and H), KitL2 (E–G), or the AP-tagged ADAM10 substrate BTC (I–K) and stimulated with 25 ng/mL PMA to activate ADAM17-dependent shedding (A, B, E, F, and H) or 2.5 µM ionomycin (IO) to activate ADAM10-dependent shedding (G and I–K) with or without siRNA against iRhom1 (siR1) (10 nM) (A, B, E, F, I, and J) or the ADAM17-specific inhibitor SP26 (G, H, and K). (C) ADAM17 Western blot shows a decrease of mature ADAM17 only in iRhom2−/− mEFs treated with iRhom1 siRNA but not in WT controls treated with iRhom1 siRNA. (D) Densitometric quantification of the percentage of mature ADAM17 relative to the proform in blots from three separate experiments like the one shown in C. *P ≤ 0.05; ±SEM (n = 3 for A–K).