Figure 3: Characterization of β-cell apoptosis and autoimmune responses in SENP1-deficient mice. | Nature Communications

Figure 3: Characterization of β-cell apoptosis and autoimmune responses in SENP1-deficient mice.

From: SENP1-mediated NEMO deSUMOylation in adipocytes limits inflammatory responses and type-1 diabetes progression

Figure 3

(a) Haematoxylin and eosin staining for overall morphology of the pancreases from Ctrl and SENP1-aP2KO mice was compared. Representative images of the pancreases from Ctrl (n=6, male) and SENP1-aP2KO mice (n=6, male) at the ages of 7, 12 and 21 weeks are shown. Three sections from each mouse. Data are representative for three independent experiments. Arrowheads indicate structural changes in the islets of SENP1-aP2KO mice. (b,c) β-Cell apoptosis. TUNEL in situ cell death staining together with α-cell marker glucagon and β-cell marker insulin. Representative images from Ctrl (n=6, male) and SENP1-aP2KO mice (n=6, male) at the age of 12 weeks are shown in b and TUNEL-positive β cells in 7, 12 and 21 weeks were quantified in c. Data are means±s.e.m. of five sections from each islet. Data are representative for three independent experiments. (d,e) CD31 and insulin co-staining from Ctrl mice (n=6, male) and SENP1-aP2KO mice (n=6, male) (d) and insulin level were calculated by Image J software (e). (f,g) T-cell infiltration. Representative images are anti-CD8 staining (red) of Ctrl (n=6, male)and SENP1-aP2KO mice (n=6, male) at the ages of 7 and 12 weeks (f) and numbers of CD8+ T cells invading into the islet of Ctrl and SENP1-aP2KO mice were quantified (g). Data are means±s.e.m. of five sections from each islet. Data are representative for three independent experiments. (h–j) Autoimmune markers. Insulin autoantibody (IAA), CRP and β-hydroxybutyrate were determined using ELISA. (k) Number of macrophages, T cells and B cells in pancreases were measured and quantified by FACS at the age of 12 weeks. (l) T-cell subset distribution. Percentages of Th1 (IFN-γ+CD4+) effector T cells, Th17 (IL-17 A+CD4+) effector T cells and regulatory T cells (CD4+CD25+Foxp3+) in pancreatic lymph nodes of Ctrl and SENP1-aP2KO mice were detected by FACS and quantified. (m) Percentage of DCs in pancreatic lymph nodes was quantified by FACS at the age of 12 weeks. (n,o) CD8+ T cells isolated from pancreatic lymph nodes (n) and spleen (o) of 12-week-old Ctrl and aP2KO mice were stained with TUM (Ctrl) and islet autoantigen (NRP-V7)-specific tetramers followed by quantifications by FACS. Scale bar, 20 μm (in all images). The two-tailed Student’s paired t-test was used for the statistical analysis. All data are presented as means±s.e.m. from six paired samples of Ctrl and SENP1-aP2KO male mice at each age. *P<0.05; **P<0.01; ***P<0.001. Ctrl, control; DS, dendritic cell; KO, knockout; LN, lymph node.

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