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: Journal of tissue science & engineering

Article Title: Whole-Cell Dissociated Suspension Analysis in Human Brain Neurodegenerative Disease: A Pilot Study

doi:

Figure Lengend Snippet: Inorganic and organic supplements.

Article Snippet: , Mouse anti-GFAP (Clone 1B4) Alexa Fluor 647 , Cow spinal cord homogenate , BD Pharmingen/560298.

Techniques: Marker, Recombinant, Purification, Residue, Sequencing