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Fig. 8. Morphological and functional analysis of Brm^-/-/Brg1^ep-/-(ib) mutant fetuses. (A) Gross morphology of (a) Brm^+/-/Brg1^L2/L2 and (b) Brm^-/-/Brg1^ep-/-(ib) E18.5 fetuses. (B) Histological analysis of Toluidine Blue-stained semithin sections of dorsal skin biopsies from E18.5 Brm^+/-/Brg1^L2/L2 (a) and Brm^-/-/Brg1^ep-/-(ib) (b) fetuses. Epidermal cell layers are indicated: C, cornified; G, granular; S, spinous; B, basal; D, dermis. Scale bar in a: 12 µm. (C) Immunohistochemical detection of proliferation marker KI67 (pink colour) on dorsal skin sections from E18.5 control (a), Brg1^ep-/-(c) (b) and Brm^-/-/Brg1^ep-/-(ib) (c) fetuses. Nuclei are stained with DAPI (blue). B, basal; SB, suprabasal; D, dermis. Scale bar in a: 25 µm. The dotted line indicates the dermal-epidermal junction. (D) Percentage of KI67-positive basal cells in the dorsal skin of E18.5 control, Brg1^ep-/-(c) and Brm^-/-/Brg1^ep-/-(ib) fetuses. The number of KI67-positive cells out of 3000 DAPI-stained interfollicular basal cells were determined on four 10-µm thick frozen skin sections from three controls and three mutants. Values expressed as a percentage are the mean values±s.e.m. (n=12). (E) Transmission electron microscopy of dorsal skin from E18.5 Brm^+/-/Brg1^L2/L2 (a) and Brm^-/-/Brg1^ep-/-(ib) (b) fetuses. SC, stratum corneum; SG, stratum granulosum; SS, stratum spinosum. Arrows point to swollen cells in the spinous and granular layers. Scale bar: 4 µm. (F) Barrier-dependent X-Gal diffusion assay on E18.5 Brm^+/-/Brg1^L2/L2 (a) and Brm^-/-/Brg1^ep-/-(ib) (b) fetuses. (G) TEWL on dorsal and ventral skin of E18.5 Brg1^L2/L2, Brg1^ep-/-(ib), Brm^+/-/Brg1^L2/L2 and Brm^-/-/Brg1^ep-/-(ib) fetuses. ^*P<0.05.

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