Extended Data Fig. 4 |. Loss of iRHOM2 in lung epithelial cells affects ADAM17 maturation/activity.

(a) Flow cytometry dot plots confirming the enrichment of mouse non-hematopoietic CD326⁺ epithelial cells isolated from total lung cells from two wild type (WT) or Rhbdf2^−/− (KO) mice by magnetic separation. CD45, hematopoietic cell marker; CD326, epithelial cell marker. (b) Western blot as in Fig. 4 g, of A549 lung tumor cells treated with the indicated siRNA. (c) Q-PCR of RHBDF1, RHBDF2 and ADAM17 in A549 cells treated with the indicated siRNAs or control RNA for 48 hours. RQ, relative quantitation. Data of three independent experiments (n = 3 per group). (d) Q-PCR as in (c), in A549 cells (n = 3 per group). (e) Q-PCR as in (c), in H1299 cells (n = 3 per group). (f) Q-PCR as in (c), in PC9 cells (n = 3 per group). (g) Q-PCR of AREG, HBEGF and TGFA in A549 cells knocked out of indicated targets (n = 3 per group). (h) Wound healing assay as in Fig. 4j, in knockout of indicated targets in A549 cells (n = 3 per group). (i) Wound healing assay as in Fig. 4j, in knockout H1299 cells either with or without HB-EGF (n = 9 per group). (j) Wound healing assay as in Fig. 4i, in knockout A549 cells (n = 9 per group). (k) Wound healing assay as in Fig. 4i, in knockout PC9 cells (n = 9 per group). All data are mean ± s.d. and were analyzed by two-tailed, unpaired Student’s t-test (h-k).