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Fig. 4 | Arthritis Research & Therapy

Fig. 4

From: Conditional deletion of caspase-8 in macrophages alters macrophage activation in a RIPK-dependent manner

Fig. 4

RIPK3 deletion prevents inflammation in Cre LysM Casp8 fl/fl mice. 2–3-month-old (young) and 6–8-month-old (aged) female Casp8 fl/fl (control), Cre LysM Casp8 fl/fl, and RIPK3 −/− Cre LysM Casp8 fl/fl mice (n ≥ 4) were evaluated for systemic autoimmune disease phenotypes, and splenocytes from aged mice were analyzed by flow cytometry. a Spleen weights of young and aged mice. b Total numbers of live splenocytes from aged mice. c Cervical lymph node weights of young and aged mice. d Formalin-fixed kidney sections (5 μm) from aged mice stained with periodic acid–Schiff (PAS). e Kidney scores of aged mice. f Proteinuria of aged mice assessed using Uristix reagent strips. Serum from aged mice was evaluated for levels of g ssDNA-, dsDNA-, histone-, and chromatin-reactive IgG antibodies and h cytokines and chemokines. i Number of CD11b+F4/80Ly6G+ neutrophils, Ly6Chigh and Ly6Clow CD11b+F4/80+ cells and CD11bF4/80+ red pulp macrophages. j Number of CD11c+CD8 and CD11c+CD8 conventional DCs and CD11cintermediatePDCA-1+B220+ plasmacytoid DCs. k Total B-cell (CD11cB220+) numbers. l B-cell subsets: follicular (FO; CD19+CD21/35+CD23+), marginal zone (MZ; CD19+CD21/35+CD23low), transitional 1 (T1; B220+AA4.1+CD23), transitional 2 (T2; B220+AA4.1+CD23+), plasmablasts (PB; CD19+B220lowCD138+CD21/35CD23), and plasma cells (PC; CD19+B220+CD138+CD21/35CD23). m Total CD4+ and CD8+ T-cell numbers. Naïve (CD44CD62L+), central memory (CD44+CD62L+), and activated (CD44+CD62L) n CD4+ and o CD8+ T-cell numbers. p CD4CD8CD3+B220+ double-negative T-cell numbers. q CD4+CD25+Foxp3+ regulatory T-cell numbers. ah represent analyses that included data from control and Cre LysM Casp8 fl/fl mice shown in Figs. 1b–d, f–h, and j. Data are represented as mean ± SD and were compared by Mann–Whitney U test: *p < 0.05; **p < 0.005; ***p < 0.0005. Casp8 caspase-8, LPS lipopolysaccharide, IL interleukin, Gro-α growth-regulated oncogene-α, IFN interferon, Ig immunoglobulin, KC keratinocyte chemoattractant, TNF tumor necrosis factor, MCP monocyte chemoattractant protein, LN, lymph node, OD optical density, RIPK receptor-interacting serine/threonine protein kinase, PDCA-1 plasmacytoid dendritic cell antigen 1, sRANKL soluble receptor activator of nuclear factor κB ligand

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