Fig 1: PTX3 is increased in tau-/- astrocytic culture. (A) Representative images of WT and tau-/- astrocyte cultures treated with 1 µM Aß oligomers, stained for PTX3 and GFAP. ns astrocytes were used as a control. Bar = 20 µm (B) Quantification of the mean fluorescence intensity of PTX3 in the conditions shown in A. n = 3, Shapiro–Wilk normality test, one-way ANOVA test, significance = P < 0.05. P values indicated on the graph. Data are presented as the mean ± SEM. (C) PTX3 concentration in astrocyte cultured media of ns or Aß-stimulated WT and tau-/- astrocyte cultures. n = 3 Shapiro–Wilk normality test, one-way ANOVA test, significance = P < 0.05. P values indicated on the graph. Data are presented as the mean ± SEM.
Fig 2: Tau-/- astrocytes exhibit a protective-type nature. (A) Volcano plot of the differential gene expression between WT and tau-/- astrocytes in the Glial Profiling Panel from Nanostring®. The analysis was performed using nSolver® software. The false discovery rate corrected level of significance is shown as the horizontal line. Significance was set at P < 0.05 and log2 Fold Change (vertical lines) at -1 & 1. (B) Undirected Global Significance Scores for the top nine gene set annotations provided by nSolver® software from the comparison between WT and tau-/- astrocytes. (C) Violin plot representation of the A2 gene set annotation score in WT or tau-/- astrocytes. Shapiro–Wilk normality test, Student’s t-test, P < 0.05. (D) Gene expression heatmap of differentially expressed A2 annotation-related genes between WT and tau-/- astrocytes. (E) & (F) Normalized counts of PTX3 and Cd109 mRNA. Shapiro–Wilk normality test, Student’s t-test, P < 0.05. n = 3 for all experiments. Data on C, E and F are presented as the mean ± SEM.
Fig 3: PTX3 rescues the synaptotoxic effects of Aß stimulated ACM. (A) Schematic representation of the methodology used in the present experiments. (B) Representative images of 14 DIV WT neurons treated with 10 µg of total protein from ns or Aß-stimulated (+Aß) ACM with or without 1 µg/ml recombinant PTX3 or heat-inactivated PTX3 (iPTX3). (C–E) After 24 h of incubation, the numbers of Synapsin-1, PSD95 and synaptic clusters were quantified. n = 4 cultures. Shapiro–Wilk normality test, one-way ANOVA, significance = P < 0.05. P values shown on each graph. Data is presented as the mean ± SEM. Bar = 10 µm.
Fig 4: Astrocytic tau expression silencing via short hairpin increases astrocyte-derived PTX3 levels. (A) Schematic representation of the methodology used in the present experiments. (B) Representative western blot for total tau in shScr or shTau transduced astrocytes lysates. ß-Actin was used as the loading control. See Supplementary material for uncropped blot. (C) Representative images of WT shScr or shTau transduced astrocyte cultures treated with 1 µM Aß oligomers stained for PTX3 and GFAP. ns astrocytes were used as the control. Bar = 20 µm. (D) Quantification of the mean fluorescence intensity of PTX3 on the conditions shown in C. All experiments n = 3. Shapiro–Wilk normality test, one-way ANOVA test, significance = P < 0.05. P values indicated on the graph. Data are presented as the mean ± SEM.
Fig 5: PTX3 is increased in the tau-/- mouse brain. (A) Representative images of the cortex from 6-month-old WT and tau-/- mice stained for PTX3 and GFAP. Bar = 50 µm on 20X and 20 µm on 60X. (B) Quantification of the colocalization index of PTX3 and GFAP. n = 6 Shapiro–Wilk normality test, one-way ANOVA test, significance = P < 0.05. P values indicated on the graph. Data are presented as the mean ± SEM.
Supplier Page from Abcam for Mouse PTX3 ELISA Kit