Fig. 2

Loss of Krt76 leads to enlarged lymph nodes and spleen, without affecting tongue and stomach epithelial homoeostasis. Immunostaining (a) and quantification (b) of EdU-labelled cells per mm of tongue and squamous stomach epithelia (n = 3 mice/genotype, 2 sections/mouse and >6 fields quantified per section, means ± s.e.m. are shown). c Hematoxylin-eosin (H&E) stained sections. d Krt76^−/− 6 month-old mouse with a neck cyst (arrowed). e Mean age (±s.e.m.) of onset of macroscopic neck lymphoid cysts in Krt76^−/− (n = 16) and control mice (n = 41) (****p ≤ 0.0001, unpaired t-test). f Macroscopic views of submandibular lymph nodes from control and Krt76^−/− mice and cyst from Krt76^−/− mouse. Pictures are representative of 9 mice/genotype. g Hematoxylin-eosin stained sections of neck lymphoid cyst inside the lymph nodes (LN) at early (left) and advanced (right) stages, juxtaposed to the salivary glands (SG). h, i Immunofluorescence staining of lymph nodes and lymphoid cyst sections with (h) anti-B220 (B cell marker) and (i) anti-CD3 (T cell marker) with DAPI counterstain. j Representative flow cytometry dot plots showing T cell populations from thymus (CD4⁺ and CD8⁺), submandibular lymph nodes (CD4⁺ or CD8⁺) and neck lymphoid cyst (CD4⁺ or CD8⁺, as the lymph nodes) from Krt76^−/− and control mice, showing percentages of total live cells (n = 3 experiments, n = 4 mice/experiment/genotype). k Lymph nodes from control and Krt76^−/− mice. Pictures are representative of 9 mice from each genotype. l Quantification of the total absolute number of cells in lymph nodes of control and Krt76^−/− mice. m, n Thymus size (m) and thymus cell number (n) in control and Krt76^−/− mice. o Representative photograph of thymus from control and Krt76^−/− mice. p, q Spleen weight (p) and absolute spleen cell numbers (q) in control and Krt76^−/− mice. r Representative photograph of spleens from control and Krt76^−/− mice. s Total body weight of control (n = 27) and Krt76^−/− (n = 10) mice. l, m, n, p, q mean ± s.e.m., *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001, unpaired t-test, n = 4 mice/genotype, measured in duplicate and experiment repeated twice. Scale bars: 100 µm (a, c, g–i), 500 µm (f, k, o, r). Sub submandibular, Ax axillar, Mes mesenteric, Ing inguinal