Fig. 2

Scrib KO embryos display normal developmental timing, proliferation and tight junction distribution. a Analysis of forelimb skeletal morphology following alcian blue and alizarin red staining at E17.5. No obvious developmental delay was observed in Scrib KO forelimbs when compared with Wt or Het littermates (scale bar = 2 mm, n = 3–6). b At E13.5, the characteristic pattern of interdigital apoptosis as revealed by Lysotracker incorporation (white arrowheads) indicates no obvious developmental delay in Scrib KO embryos when compared with Wt or Het littermates (scale bar = 1 mm, n = 3). c Whole E17.5 embryo skeletal analysis (scale bar: 5 mm, n = 3–6) reveals extensive dysmorphology in Scrib KO embryos, including (d) loss of dorsal cranial bones such that the basi-occipital bone (asterix) visible in dorsal view, (e) vertebral fusion (e, red arrowhead), hemivertebra (e, yellow arrowhead) and (f) distal rib fusion . Scale bar in d-f = 1 mm, n = 3. g IHC to detect PCNA in Scrib Wt, Het and KO embryonic epidermis at E16.5 (scale bar: 50 μm, n = 3, dashed line represents basement membrane). h IF images to detect ZO-1 (green) in Scrib Wt, Het and KO embryonic epidermis at E17.5 (scale bar = 50 μm, DAPI = blue, n = 3, dashed line represents basement membrane). i Tight junction ultrastructure analysis within the granular layer of Scrib Wt, Het and KO embryonic epidermis at E17.5 at 8,000x, 33,000x and 46,000x magnification (scale bar = 0.5 μm, TJ = tight junction and D = desmosome, n = 3–6). Scrib Wt, Het and KO embryos displayed a similar frequency of TJs with normal morphology within the SG