Fig 1: Differential relative abundance of top serum proteins in pre-diagnostic PM cases versus controls. Box plots show relative LC-MS/MS intensities for each DEP grouped by time to diagnosis (0–2 years, 2–5 years) and matched controls. Up-regulated proteins include Complement C4A (C4; FC = 1.4, p = 0.009) and Transferrin (TF; FC = 1.4, p = 0.008). Down-regulated proteins comprise Beta-2-Microglobulin (B2M; FC = 0.7, p = 0.035), and Dermcidin (DCD; FC = 0.6, p = 0.029). Asterisks indicate nominal p < 0.05 by GLM
Fig 2: Proteomic analysis reveals consistent reductions in neuronal, synaptic, and oligodendroglial proteins in monocyte-engrafted mice.(a) Heatmap showing log2-transformed intensities in a subset of 74 non-myeloid DEPs between microglia and monocyte engrafted brains (n=8/group). (b) ELISA quantification of Synaptophysin (SYP), Post Synaptic Density-95 (PSD-95), Myelin Basic Protein (MBP), Glial Fibrillary Acidic Protein (GFAP), Complement component 4 (C4), Tumor Necrosis Factor-Alpha (TNF-α), Interleukin-1 beta (IL-1β), Interleukin-2 (IL-2), Interleukin-8 (IL-8), Interleukin-12 (IL-12), Interleukin-34 (IL-34), and Interferon gamma (IFN-γ) in soluble half-brain lysates; p values from one-way ANOVA (SYP: F(2,28)= 24.65; ****p<0.0001) (PSD-95: F(2,28)= 3.762; *p=0.0357) (MBP: F(2,28)= 27.82; ****p<0.0001) (GFAP: F(2,28)= 18.76; ****p<0.0001) (C4a: F(2,28)= 26.63; ****p<0.0001) (TNF-α: F(2,28)= 7.276; **p=0.0029) (IL-1β: F(2,28)= 77.13; ****p<0.0001) (IL-2: F(2,28)= 5.828; **p=0.0077) (IL-8: F(2,28)= 9.766; ***p=0.0006) (IL-12: F(2,28)= 22.34; ****p<0.0001) (IL-34: F(2,28)= 111.7; ****p<0.0001) (IFN-γ: F(2,28)= 7.455; **p=0.0025) with Tukey multiple comparisons tests. Data represented as average mean value ± SEM (WT, n = 15; Mg, n=8; Mo, n = 8). (c) Matrix of Pearson correlation (r) coefficients of quantified SYP, PSD-95, MBP, GFAP, C4, TNF-α, IL-1β, IL-2, IL-8, IL-12, IL-34, and IFN-γ. (d-e) Simple linear regression between levels of SYP and C4 (R2 = 0.719; ****P<0.0001), SYP and TNF-α (R2 = 0.577; ***P=0.0006), MBP and C4a (R2 = 0.558; ***P=0.0009), and MBP and TNF-α (R2 = 0.4348; **P=0.0055) in xenografted mice. (f) Representative confocal imaging of neuronal nuclei (NeuN, green) and xenografted microglia or monocytes (IBA1, green) in the ventral dentate gyrus, scale bar: 40μm. (g) Quantification of NeuN integrated density within the upper blade of the dentate gyrus of control (WT) or Monocyte (Mo) and Microglia (Mg) engrafted mice. As group distributions of NeuN deviated from normality, we utilized a non-parametric Kruskal–Wallis test (P=0.042) followed by Dunn’s multiple comparisons test, monocyte versus WT mice (Dunn’s: P=0.049), microglia versus WT mice (Dunn’s: P>0.999). (h) Representative confocal imaging of a neuronal nuclei (NeuN, green) surrounded by phagocytic (CD68, yellow) monocytes (IBA1, green) within the dentate gyrus, scale bar: 10μm. Data represented as average mean value ± SEM (WT, n = 8; Mg, n=6–7; Mo, n = 11). *p < 0.05, **p < 0.01, ***p < 0.0001, and ****p < 0.0001. Comparisons not shown are not significant.
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