FIGURE 3.

Cdh16‐Cre/Mfn2 ^flox/flox mice develops malignant lesions and exhibits EGFR signaling pathway activation. (A) Experimental scheme for generating the Mfn2 ^CKO mice. (B) Representative images of kidneys of Mfn2 ^CKO and control Mfn2 ^flox/flox mice taken at 40 weeks of age. (C) Representative H&E staining of kidney sections of 40‐week‐old control and Mfn2 ^CKO mice. (D) Representative H&E staining of kidney sections. (i) H&E staining of control mice display normal kidney morphology. (ii‐vi) H&E staining of kidney sections of Mfn2 ^CKO mice shows solid dysplasia (ii), clear cytoplasm (iii), a simple cyst lined by a single epithelial layer (iv), a cyst showing atypical epithelial cell proliferation (multi‐layered) (v) and a cyst showing extensive proliferation of epithelial cells into the cyst lumen (vi). (E) Representative IHC staining images of p‐EGFR, p‐AKT and p‐ERK in kidney sections of control and Mfn2 ^CKO mice (n = 3 per group). IHC scores are shown. (F) Western blotting analysis of indicated proteins in EGF‐treated primary renal tubular epithelial cells isolated from control and Mfn2 ^CKO mice. (G) Representative H&E staining of kidney sections from Ctrl, Vhl ^CKO, Mfn2 ^CKO and VhlMfn2 ^CKO mice.

Abbreviations: Mfn2 ^CKO, Cdh16‐Cre/Mfn2 ^flox/flox mice; CKO, conditional knockout; Ctrl, control Mfn2 ^flox/flox mice; H&E, hematoxylin and eosin; EGF, epidermal growth factor; Vhl, Von Hippel‐Lindau Tumor Suppressor; Vhl ^CKO, Cdh16‐CreVhl ^flox/flox mouse; VhlMfn2 ^CKO, Cdh16‐Cre/Vhl ^flox/flox /Mfn2 ^flox/flox mice.