Journal: iScience

Article Title: Type 2 and type 1 diabetes have opposing effects on the systemic murine complement alternative pathway

doi: 10.1016/j.isci.2026.116359

Figure Lengend Snippet: HFD leads to downregulation of the expression of adipose Cfd and hepatic C3 and Cfb (A and B) C3-tdTomato reporter mice were placed on LFD or HFD, and expression of inflammatory markers and complement genes were assessed by qPCR in (A) subcutaneous inguinal adipose tissue and (B) visceral epididymal adipose tissue. (C) Western blot for FD in serum from female LFD and HFD mice after 25 weeks of diet. (D) Western blot for FD in serum from male mice after 15 weeks of diet. (E) Left: representative western blot from sequential serum samples from 2 male mice on HFD. Right: densitometry for serum FD signal in western blots of 6 male mice over time. (F–H) Serum FD levels, measured by ELISA, plotted against individual mouse body weight (F), fasting insulin (G), and fasting blood glucose levels (H). (I) Expression of main alternative pathway components in livers of LFD and HFD mice. (J) Protein levels of C3 in liver homogenates from lean control or ob/ob mice on the C57Bl/6 background. WAT, white adipose tissue. In (A)–(C), there were 4 mice per group, except for HFD visceral fat pad, which had 3 samples. In (F)–(H), measurements are from a total of 42 individual mouse samples. For (I) and (J), there were 4–5 mice per group. Data represent mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, as tested by two-way ANOVA. See also .

Article Snippet: Serum insulin was measured using an ultrasensitive mouse insulin ELISA (Mercodia #10-1249) and mouse C3b was measured in serum samples using an ELISA kit from Hycult Biotech (HK216).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: iScience

Article Title: Type 2 and type 1 diabetes have opposing effects on the systemic murine complement alternative pathway

doi: 10.1016/j.isci.2026.116359

Figure Lengend Snippet: Expression changes in STZ-induced diabetes leads to increased complement in serum and increased alternative pathway activation (A) Serum levels of FD in untreated and STZ-treated male mice. (B) Serum C3 levels in untreated controls and STZ-treated male mice. (C) Western blot for FD in homogenates of inguinal adipose tissue from control mice and STZ-treated WT and C3-KO mice. Densitometry quantification shown on the right. (D) Blood glucose levels in STZ-treated WT or C3-KO male mice over time. (E) Serum FB levels in STZ-treated male mice before and after treatment. (F) Western blot for FB in serum samples of untreated and STZ-treated male mice (top), and quantification by densitometry (bottom). (G) Example flow cytometry histograms of C3 staining of zymosan beads after incubation with serum from untreated or STZ-treated cage-mate mice in EGTA buffer, allowing only AP activation. EDTA completely inhibits complement and acts as a negative control. (H) Results of C3 deposition onto zymosan beads from serum taken at different time points from untreated or STZ-treated male mice. (I) Serum C3b levels in STZ-treated or mock-treated male mice and in STZ-treated C3-KO controls, as measured by ELISA. For all images, n = 4 untreated mice and 5 STZ-treated mice, as in , except for (C), with groups of 4; (D), with groups as stated in the figure; and (I), with n = 8, 9, and 4 for untreated, STZ-treated, and STZ-treated KO groups, respectively. Data represent mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, as tested by two-way ANOVA, t test (E), or one-way ANOVA (C and I).

Article Snippet: Serum insulin was measured using an ultrasensitive mouse insulin ELISA (Mercodia #10-1249) and mouse C3b was measured in serum samples using an ELISA kit from Hycult Biotech (HK216).

Techniques: Expressing, Activation Assay, Western Blot, Control, Flow Cytometry, Staining, Incubation, Negative Control, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Type 2 and type 1 diabetes have opposing effects on the systemic murine complement alternative pathway

doi: 10.1016/j.isci.2026.116359

Figure Lengend Snippet: T1D Akita mice have increased serum FB levels and alternative pathway activation (A) Serum FD levels in Akita and WT littermates, as measured by ELISA. (B) Serum C3 levels in the same mice. (C) Serum C3 levels in “young” versus “older” mice. (D) Serum FB levels in all Akita and WT littermate mice. (E) Serum FB levels in the same mice, stratified over time, plotting line of best fit and 95% confidence intervals. (F) C3 alternative pathway deposition onto zymosan beads from age-matched Akita or WT littermate mice. Left: example histograms from cage-mate mice; right: quantification of C3 deposition results from serum taken from age-matched pairs of mice aged from 18 to 24 weeks ( n = 5 per genotype). Each data point represents mean value from an individual mouse. Data represent mean ± SD, with ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, as tested by one-way ANOVA (F) or t test (C and D). In (E), lines show best fit and 95% confidence intervals.

Article Snippet: Serum insulin was measured using an ultrasensitive mouse insulin ELISA (Mercodia #10-1249) and mouse C3b was measured in serum samples using an ELISA kit from Hycult Biotech (HK216).

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay