Figure 2: Identification and correction of pathological phenotypes of DS astroglia. | Nature Communications

Figure 2: Identification and correction of pathological phenotypes of DS astroglia.

From: Role of astroglia in Down’s syndrome revealed by patient-derived human-induced pluripotent stem cells

Figure 2

(a1–6) qPCR analysis of S100B, GFAP, iNOS, TSP-1, TSP-2 and NFE2L2 mRNA expression in DS and control (Cont) astroglia. (a7) Quantification of nitrite/nitrate concentration in ACM collected from DS and Cont astroglia. One-way analysis of variance (ANOVA) test, ♣P<0.05, ♣♣P<0.01 and ♣♣♣P<0.001, comparison between two DS astroglia with Cont 1 astroglia. #P<0.05, ##P<0.01 and ###P<0.001, comparison between two DS astroglia with Cont 2 astroglia. Student’s t-test, *P<0.05, *P<0.01 and *♣P<0.001, comparison between two DS astroglia or two Cont astroglia. n=3–4 for each cell line. (b) Representative and quantification of ROS production in Cont and DS astroglia. Green fluorescence marks cells that undergo oxidation. Student’s t-test, **P<0.01. n=3–4 from each cell line. (c) Quantification of pooled data showing the proliferation rate of Cont and DS astroglia. Student’s t-test, **P<0.01. n=3–4 from each cell line. (d) Glutamate uptake analysis showing that both DS and Cont astroglia were capable of glutamate uptake. Notice that DS astroglia show glutamate uptake at a higher rate at the 30- and 60-min time point than Cont astroglia. Student’s t-test, **P<0.01 and ***P<0.001, n=4 from each cell line. (e) Representatives showing intracellular uptake of the BLOCK-iT Fluorescent Oligo at 24 h after transfection of DS astroglia. Scale bar, 50 μm. (f) qPCR analysis of pooled data showing the expression of S100B gene in DS astroglia at 48 h after transfection with Cont and S100B siRNA. Student’s t-test, **P<0.01, n=3–5 from each cell line. (g) Representatives and quantification of pooled data showing ROS production in DS astroglia determined at 48 h after transfection with Cont and S100B siRNA. Student’s t-test, *P<0.05, n=5 from each cell line. (h,i) Quantification of pooled data showing the concentration of nitrite/nitrate in the ACM and proliferation rate of DS astroglia at 48 h after transfection with Cont and S100B siRNA. Student’s t-test, *P<0.05, n=3–4 from each cell line. (j) qPCR analysis of S100B, iNOS and NEF2L2 mRNA expression in DS1 astroglia after the treatment of resveratrol, cucurmin or minocycline for 72 h. One-way ANOVA test, *P<0.05, **P<0.01 and ***P<0.001; n=3–5 for each group. (k) Quantitative analysis of the proliferation rate of DS1 and 2 astroglia after the treatment of minocycline. Student’s t-test, *P<0.05, n=3–4 for each cell line. (l) qPCR analysis of GFAP, TSP-1 and TSP-2 mRNA expression in DS astroglia after the treatment of minocycline. Student’s t-test, *P<0.05 and **P<0.01; n=3–4 for each group. Data are presented as mean±s.e.m.

Back to article page