Desiccating stress-induced changes in wild-type (WT) and DNTGFβRII mice at 14 weeks of age (14W).

<p>Desiccating stress was induced for 5 days in both strains (DS5); nonstressed mice served as controls (NS). <b>A.</b> Representative images of OGD corneal staining used to generate OGD intensity score in <b>B.</b> Bar charts show mean ± SD of three independent experiments (final n = 12 for each group). <b>C.</b> CD4 counts in corneal epithelium (EPI) and corneal stroma (ST). Bar charts show mean ± SD of three independent experiments (final n = 5 for each group). <b>D.</b> Representative images of conjunctiva sections immunostained for CD4 (in red) used to generate CD4 counts in <b>E</b>, in conjunctiva epithelium Bar charts show mean ± SD of three independent experiments (final n = 5 for each group). <b>F.</b> PAS+ conjunctival goblet cells counts. Bar charts show mean ± SD of three independent experiments (final n = 5 for each group). <b>G–I.</b> Relative fold of expression in cornea (I), conjunctiva, (CJ, in J) and lacrimal gland (LG, in K) using the WT-NS of each strain as the strain-calibrator. Bar charts show mean ± SD of three independent experiments (final n = 8 for each group). **P<0.01;*** P<0.001 indicate within WT comparison (NS vs. DS5) ∧ P<0.05;∧∧∧P<0.01 indicates DNTGFBRII comparison (NS vs. DS5).</p

Expression of chemokine receptors and adoptive transfer results in RAG1KO mice.

<p><b>A–B</b> Flow cytometry analysis of freshly isolated cells from ocular surface (OS, in A) and cervical lymph nodes (CLN, in B) from wild-type (WT) and DNTGFβRII mice before (nonstressed, NS) and after 5 days of desiccating stress (DS5).*P<0.05, within strain comparison. Bar charts show mean ± SD of three independent experiments (final n = 4 for each group for OS, final n = 6 for each group for CLN). <b>C.</b>Conjunctival sections stained for CD4⁺ T cells (in red) of RAG1KO recipient mice that received CD4+ cells isolated from WT and DNTGFβRII DS5 mice. Note CD4+T cell infiltration in goblet cell rich area of conjunctiva in WT-DS5 recipient mice. The black dotted circle is a higher magnification of area demarcated by blue dotted circle. Original magnification: 20×. Scale bar = 50 µm. <b>D and E</b>. Conjunctival CD4+T cell <b>(in D)</b> and PAS+ conjunctival goblet cells <b>(in E)</b> counts in RAG1KO recipient mice that received cells from CD4+ cells isolated from WT and DNTGFβRII mice after 5 days of desiccating stress (DS5). Bar charts show mean ± SD of three independent experiments (final n = 5 for each group) <b>F</b>.Relative change of IL-17A, IFN-γ and IL-13 mRNA transcripts in conjunctiva of RAG recipients. Asterisks indicate comparison to strain-specific controls. Bar charts show mean ± SD of two independent experiments (final n = 6 for each group).</p

Age-related changes in wild-type (WT) and DNTGFBRII mice, from 8 to 14 weeks of age (8W and 14W, respectively).

<p><b>A.</b> Representative images of cornea sections immunostained for CD4 (in red) used to generate CD4 counts in B, in corneal epithelium (EPI) and corneal stroma (ST) in wild-type and DNTGFβRII mice at 8 and 14 weeks of age (8W,14W). Bar charts show mean ± SD of two independent experiment (final n = 5 for each group). <b>C.</b> Representative images of OGD corneal staining used to generate OGD intensity score in <b>D.</b> Bar charts show mean ± SD of three independent experiments (final n = 12 for each group). <b>E.</b> Corneal smoothness score. Bar charts show mean ± SD of three independent experiments (final n = 13 for each group). <b>F.</b> Representative images of conjunctiva sections immunostained for CD4 (in red) used to generate CD4 counts in <b>G.</b> Bar charts show mean ± SD of three independent experiments (final n = 5 for each group). <b>H.</b> PAS+ conjunctival goblet cells counts. Bar charts show mean ± SD of three independent experiments (final n = 5 for each group). <b>I–K.</b> Relative fold of expression in cornea (I), conjunctiva, (CJ, in J) and lacrimal gland (LG, in K) using the WT-8W as the calibrator. Bar charts show mean ± SD of three independent experiment (final n = 8 for each group). In graphs B, D,E, G, H **P<0.01;*** P<0.001 indicate within strain comparison In I–K graphs,*** P<0.001 indicates interstrain comparison.</p

CD4+ T cell proliferation.

<p>Proliferation of CD4⁺ T cell in CD4⁺ T cells co-cultured in the presence of cornea and conjunctival tissues (CN and CJ, respectively) before (nonstressed [NS]) or after desiccating stress for 5 days [DS5] from wild-type (WT) and CD4-DNTGFβRII mice (DN). Bar charts show mean ± SD of two independent experiments (final n = 5 for each group).</p