Fig. 1
From: Immunomodulatory role of Keratin 76 in oral and gastric cancer
Keratin 76 is expressed in the oral epithelia and squamous stomach. a Krt76 knockout strategy. Krt76−/− mice were generated by disruption of the Krt76 gene via a knockout first allele targeting construct (reporter-tagged insertion with conditional potential). These animals have a splice acceptor-LacZ reporter gene integrated in the targeting gene, between exon 1 and 2, which allows tracing of gene expression whilst disrupting Krt76 protein expression. b X-gal staining (blue) of beta-galactosidase expressed under the control of the Krt76 promoter in the oral cavity and stomach (arrows) of Krt76+/− mouse embryos at E17.5. c Immunofluorescence labelling with anti-Krt76 (green) and anti-Krt14 (red) antibodies in the oral cavity and stomach of mouse embryos at E17.5. Bottom row: left hand panel is higher magnification view of boxed area in right hand panel. d Whole-mount X-gal staining of Krt76+/− reporter mice at post-natal day 2 (P2) shows Krt76 expression in the dorsal and lateral tongue, with partial expression in the ventral tongue. e–h X-gal staining (blue) of beta-galactosidase expressed under the control of the Krt76 promoter in tongue (e), palate (f), lip and buccal mucosa (g) and in stomach (h) of Krt76+/− adult mice. h Mouse stomach is subdivided into two major histologically distinct regions: the squamous stomach lined with a stratified squamous epithelium and the glandular stomach, separated by the limiting ridge from the stratified squamous epithelium of the squamous stomach. Krt76 expression is restricted to the squamous stomach region. i Immunofluorescence labelling with anti-Krt76 (green) and anti-Krt14 (red) antibodies of adult wild-type mouse tissues, confirming the specificity of both anti-Krt76 antibody and X-gal staining. Samples were counterstained with nuclear dye DAPI (4′,6-diamidino-2-phenylindole). Dotted line delineates basement membrane. j Krt76 mRNA qRT-PCR analysis of adult tissues, relative to Gapdh (n = 3 mice, means ± s.e.m. are shown). Scale Bars = 500 µm (b, c), 100 µm (e–i)
