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(a-q) Aged (20-month-old) male mice were randomly allocated and intraperitoneally injected with either vehicle or the C3 inhibitor (AMY-101; 1 mg/kg) weekly for 6 weeks and analyzed ( n = 9 for vehicle and n = 8 for C3 inhibitor). (a) Schematic diagram for the experimental design. (b) Body weight measurement. (c,d) Fat mass (c) and lean mass (d) were measured by EchoMRI. (e) Representative western blots with densitometry ( n = 4 for vehicle and n = 5 for C3 inhibitor). (f) A glucose tolerance test was performed. (g,h) Healthspan was analyzed through grip strength (g) and the <t>rotarod</t> <t>test</t> (h). (i-m) Serum levels of IL-1β (i), IL-6 (j), TNF-α (k), MCP-1 (l), and GM-CSF (m) were analyzed by Luminex assay. Only detected values were presented. (n-q) Flow cytometry analysis of VAT SVF. The frequencies of ATMs (n), AABs (o), CD8 T (p), and NK cells (q) were presented. (r-t) Representative C3 western blot analysis of the VAT of 24-month-old WT and FGF21-Tg (r), 24-month-old WT and PLA2G7 -/- (s), and 16-month-old SPARC f/f and SPARC f/f;CAG-Cre (t) mice ( n = 3/group). Unpaired two-tailed t-tests were performed, and exact p-values were presented. Data presented as mean±SEM.
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(a-q) Aged (20-month-old) male mice were randomly allocated and intraperitoneally injected with either vehicle or the C3 inhibitor (AMY-101; 1 mg/kg) weekly for 6 weeks and analyzed ( n = 9 for vehicle and n = 8 for C3 inhibitor). (a) Schematic diagram for the experimental design. (b) Body weight measurement. (c,d) Fat mass (c) and lean mass (d) were measured by EchoMRI. (e) Representative western blots with densitometry ( n = 4 for vehicle and n = 5 for C3 inhibitor). (f) A glucose tolerance test was performed. (g,h) Healthspan was analyzed through grip strength (g) and the rotarod test (h). (i-m) Serum levels of IL-1β (i), IL-6 (j), TNF-α (k), MCP-1 (l), and GM-CSF (m) were analyzed by Luminex assay. Only detected values were presented. (n-q) Flow cytometry analysis of VAT SVF. The frequencies of ATMs (n), AABs (o), CD8 T (p), and NK cells (q) were presented. (r-t) Representative C3 western blot analysis of the VAT of 24-month-old WT and FGF21-Tg (r), 24-month-old WT and PLA2G7 -/- (s), and 16-month-old SPARC f/f and SPARC f/f;CAG-Cre (t) mice ( n = 3/group). Unpaired two-tailed t-tests were performed, and exact p-values were presented. Data presented as mean±SEM.

Journal: bioRxiv

Article Title: Exoproteome of calorie-restricted humans identifies complement deactivation as an immunometabolic checkpoint reducing inflammaging

doi: 10.1101/2025.08.04.668533

Figure Lengend Snippet: (a-q) Aged (20-month-old) male mice were randomly allocated and intraperitoneally injected with either vehicle or the C3 inhibitor (AMY-101; 1 mg/kg) weekly for 6 weeks and analyzed ( n = 9 for vehicle and n = 8 for C3 inhibitor). (a) Schematic diagram for the experimental design. (b) Body weight measurement. (c,d) Fat mass (c) and lean mass (d) were measured by EchoMRI. (e) Representative western blots with densitometry ( n = 4 for vehicle and n = 5 for C3 inhibitor). (f) A glucose tolerance test was performed. (g,h) Healthspan was analyzed through grip strength (g) and the rotarod test (h). (i-m) Serum levels of IL-1β (i), IL-6 (j), TNF-α (k), MCP-1 (l), and GM-CSF (m) were analyzed by Luminex assay. Only detected values were presented. (n-q) Flow cytometry analysis of VAT SVF. The frequencies of ATMs (n), AABs (o), CD8 T (p), and NK cells (q) were presented. (r-t) Representative C3 western blot analysis of the VAT of 24-month-old WT and FGF21-Tg (r), 24-month-old WT and PLA2G7 -/- (s), and 16-month-old SPARC f/f and SPARC f/f;CAG-Cre (t) mice ( n = 3/group). Unpaired two-tailed t-tests were performed, and exact p-values were presented. Data presented as mean±SEM.

Article Snippet: A rotarod test was conducted using an accurotor rotarod (AccuScan Instruments).

Techniques: Injection, Western Blot, Luminex, Flow Cytometry, Two Tailed Test