Journal: Journal of Neuroinflammation

Article Title: Neuroprotective effect of astrocyte-derived IL-33 in neonatal hypoxic-ischemic brain injury

doi: 10.1186/s12974-020-01932-z

Figure Lengend Snippet: Astrocytic IL-33 is robustly upregulated after hypoxic-ischemia in neonatal mouse brains. a Western blot of IL-33 was performed using protein extracted from the mouse brain at 1, 3, 7, and 14 days after HI or sham operation. Data are mean ± SEM ( n = 6 in each group). # P < 0.01 compared to sham. b Representative images of IL-33 (green) and GFAP or Olig2 (red) labeling in the cerebral cortex of WT mice at 1 day after HI injury. No IL-33 staining was observed in Neun or Iba1 cells 1 d after HI. Scale bar, 25 μm. Nuclei were stained blue with 4,6-diamidino-2-phenylindole (DAPI). c Semi-quantitative analysis of glia or neuron-type cell contributions to the IL-33-positive cell population in vehicle group 1 day after HI. Data are mean ± SEM ( n = 6 in each group). * P < 0.05 compared to sham. d Representative IL-33 expression on astrocytes in WT mice at 1 day after HI injury. The numbers shown indicate the percentage of IL-33 expression on the indicated cell population. Data are mean ± SEM ( n = 6 in each group). * P < 0.05 compared to sham

Article Snippet: The sections were stained overnight at 4 °C with either anti-ST2 (1:200, ab25877, Abcam), anti-IL-33 (1:500, AF3626, R&D), anti-GFAP (1:500, 104805-T08, Sino Biological Inc.), anti-Olig2 (1:500, EPR2673, Abcam), anti-Iba1(1:50, 20A12.1, Sigma-Aldrich), anti-Neun (1:50, A60, Sigma-Aldrich), or anti-Ki67 (1:500, ab15580, Abcam) followed by PE- or FITC-conjugated secondary antibodies (eBioscience).

Techniques: Western Blot, Labeling, Staining, Expressing

Journal: Journal of Neuroinflammation

Article Title: Neuroprotective effect of astrocyte-derived IL-33 in neonatal hypoxic-ischemic brain injury

doi: 10.1186/s12974-020-01932-z

Figure Lengend Snippet: ST2 expression on astrocytes is elevated after HI. a Histograms showing ST2 expression on astrocytes, oligodendrocytes, microglia, and neurons in the brain 3 days after HI or sham operation. The blue and red lines represent staining in sham and HI brains, respectively, at 3 days after surgery. The gray area represents isotype control staining. b–e Percentages of ST2 expression on astrocytes ( b ), oligodendrocytes ( c ), microglia ( d ), and neurons ( e ) in sham brain and HI brain at 3 days after surgery. Data are mean ± SEM ( n = 5–7 per group), * P < 0.05 compared to sham. f Representative images of GFAP (green) and ST2 (red) labeling in the cerebral cortex 3 days after HI. Scale bar, 25 μm. Nuclei were stained blue with DAPI

Article Snippet: The sections were stained overnight at 4 °C with either anti-ST2 (1:200, ab25877, Abcam), anti-IL-33 (1:500, AF3626, R&D), anti-GFAP (1:500, 104805-T08, Sino Biological Inc.), anti-Olig2 (1:500, EPR2673, Abcam), anti-Iba1(1:50, 20A12.1, Sigma-Aldrich), anti-Neun (1:50, A60, Sigma-Aldrich), or anti-Ki67 (1:500, ab15580, Abcam) followed by PE- or FITC-conjugated secondary antibodies (eBioscience).

Techniques: Expressing, Staining, Labeling

Journal: Journal of Neuroinflammation

Article Title: Neuroprotective effect of astrocyte-derived IL-33 in neonatal hypoxic-ischemic brain injury

doi: 10.1186/s12974-020-01932-z

Figure Lengend Snippet: IL-33 promotes astrocyte survival and proliferation after HI injury. a Representative FACS analysis of ST2 + astrocytes at 24 h after 6 h OGD. Right, numbers indicate the percentage of ST2 + astrocytes at 24 h after 6 h OGD. Data are mean ± SEM ( n = 3 in each group). * P < 0.05 compared to untreated controls. b CCK-8 assay in astrocyte-enriched cultures subjected to 6 h OGD or sham conditions followed by treatment with PBS or a range of concentrations of IL-33 for another 24 h. Data are mean ± SEM ( n = 3 in each group). # P < 0.01 compared to untreated controls. c Representative images of Ki-67 (red) and GFAP (green) in the cerebral cortex of IL-33- and PBS-treated mice at 7 days post HI (left). Scale bar, 25 μm. Right, quantification of Ki-67 and GFAP dual-labeled cells at 7 days post HI ( n = 9 per group). d , e Percentages of phosphorylated AKT (pAKT) ( d ) and Ki-67 + ( e ) in astrocytes at 24 h after 6 h OGD and the culture conditions shown. Data are mean ± SEM ( n = 3 in each group). * P < 0.05, # P < 0.01. f Representative FACS analysis of cell-cycle status of astrocytes at 24 h after 6 h OGD and the culture conditions shown. Data are mean ± SEM ( n = 3 in each group). * P < 0.05

Article Snippet: The sections were stained overnight at 4 °C with either anti-ST2 (1:200, ab25877, Abcam), anti-IL-33 (1:500, AF3626, R&D), anti-GFAP (1:500, 104805-T08, Sino Biological Inc.), anti-Olig2 (1:500, EPR2673, Abcam), anti-Iba1(1:50, 20A12.1, Sigma-Aldrich), anti-Neun (1:50, A60, Sigma-Aldrich), or anti-Ki67 (1:500, ab15580, Abcam) followed by PE- or FITC-conjugated secondary antibodies (eBioscience).

Techniques: CCK-8 Assay, Labeling

Journal: Aging Cell

Article Title: The reverse transcriptase inhibitor 3TC protects against age‐related cognitive dysfunction

doi: 10.1111/acel.13798

Figure Lengend Snippet: 3TC improves cognitive function in a mouse model of tauopathy. (a) Re‐analysis of published RNA‐seq data showing TE transcript increases in the brains of 8‐month rTg4510 transgenic tauopathy mice versus littermate controls. (b) Novel Object Recognition index in littermate controls and rTg4510 mice treated with/without 3TC in the present study (quantified as time exploring the novel object vs. total object exploration, n = 6 mice/group; 3 months old). (c) Anxiety responses in the same mice as measured by time spent on open arms in the Elevated Plus Maze. (d, e) Immunoblot quantifications and images of Iba1 and GFAP. TE type comparisons: Shapiro–Wilk normality test followed by Mann–Whitney test versus mean Log2 fold‐change for all transcripts. Novel Object Recognition, Elevated Plus Maze and immunoblot analyses: ANOVA with Tukey's multiple comparison post‐hoc analysis. * p < 0.05; ** p < 0.01.

Article Snippet: Primary antibodies (all from ABclonal) included p‐IRF3 (1:1000 dilution), IRF3 (1:1000), p‐STING (1:1000), STING (1:1000), p‐cGAS (1:1000), cGAS (1:1000), IBA1 (1:750), pTBK1 (1:1000), TBK1 (1:1000), and GFAP (1:1000).

Techniques: RNA Sequencing, Transgenic Assay, Western Blot, MANN-WHITNEY, Comparison