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combination with ccl2  (R&D Systems Hematology)
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R&D Systems Hematology combination with ccl2
(A-C) Chemokine secretion in microglia and astrocyte supernatants, 6h after IL-1β challenge (2.5 ng/mL) measured by ELISA for <t>CCL2,</t> CXCL1 and CXCL10. Two way ANOVA, *p<0.001, compared to astrocyte control; #p<0.001, compared to microglia IL-1β. (D) Light microscopy labelling of CCL2 in WT and Tg mice, 2h after i.c. challenge with IL-1β (10 ng) or saline. (E) Confocal imaging of Tg+IL-1β group for CCL2 (red; 594nm) and GFAP (green; 488nm). Iba-1 (green; 488nm) is captured in the inset. (F) Light microscopy labelling of CXCL1. (G) Light microscopy labelling of CXCL10. Insets show detail of positive vascular and astrocytic cells for the different chemokines.
Combination With Ccl2, supplied by R&D Systems Hematology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A-C) Chemokine secretion in microglia and astrocyte supernatants, 6h after IL-1β challenge (2.5 ng/mL) measured by ELISA for CCL2, CXCL1 and CXCL10. Two way ANOVA, *p<0.001, compared to astrocyte control; #p<0.001, compared to microglia IL-1β. (D) Light microscopy labelling of CCL2 in WT and Tg mice, 2h after i.c. challenge with IL-1β (10 ng) or saline. (E) Confocal imaging of Tg+IL-1β group for CCL2 (red; 594nm) and GFAP (green; 488nm). Iba-1 (green; 488nm) is captured in the inset. (F) Light microscopy labelling of CXCL1. (G) Light microscopy labelling of CXCL10. Insets show detail of positive vascular and astrocytic cells for the different chemokines.

Journal: bioRxiv

Article Title: Microglial and Astrocyte priming in the APP/PS1 model of Alzheimer’s Disease: increased vulnerability to acute inflammation and cognitive deficits

doi: 10.1101/344218

Figure Lengend Snippet: (A-C) Chemokine secretion in microglia and astrocyte supernatants, 6h after IL-1β challenge (2.5 ng/mL) measured by ELISA for CCL2, CXCL1 and CXCL10. Two way ANOVA, *p<0.001, compared to astrocyte control; #p<0.001, compared to microglia IL-1β. (D) Light microscopy labelling of CCL2 in WT and Tg mice, 2h after i.c. challenge with IL-1β (10 ng) or saline. (E) Confocal imaging of Tg+IL-1β group for CCL2 (red; 594nm) and GFAP (green; 488nm). Iba-1 (green; 488nm) is captured in the inset. (F) Light microscopy labelling of CXCL1. (G) Light microscopy labelling of CXCL10. Insets show detail of positive vascular and astrocytic cells for the different chemokines.

Article Snippet: For confocal microscopy, sections were labelled with the following combinations: 6E10 (1:1000; Biolegend 803001) with an Alexa Fluor 488 anti-mouse (1:800; Invitrogen) together with Iba-1 (1:2000; Abcam ab5076) Alexa Fluor 594 anti-goat (1:800; Invitrogen); 6E10 with an Alexa Fluor 633 anti-mouse (1:800; Invitrogen) in combination with IL-1β (1:50; Peprotech 500-P51) with an Alexa Fluor 488 anti-rabbit (1:800; Invitrogen); 6E10 with an Alexa Fluor 633 anti-mouse and GFAP with an Alexa Fluor 488 anti-rabbit; GFAP (1:2000; Dako Z0334) with an Alexa Fluor 594 anti-rabbit (1:800; Invitrogen) together with IL-1β (1:50; Peprotech 500-P51) with an Alexa Fluor 488 anti-rabbit; GFAP with an Alexa Fluor 488 anti-rabbit in combination with CCL2 (1:200; R&D AF-479-NA) with Alexa Fluor 594 anti-goat; Iba-1 (1:2000; Abcam ab5076) with Alexa Fluor 594 anti-goat.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Light Microscopy, Saline, Imaging

Exogenous i.c. administration of LPS in normal mice induces activation of microglial cells to produce regulated secretion of IL-1β and this IL-1β may activate astrocytes to release chemokines such as CCL2, CXCL1 or CXCL10. However, in APP/PS1 mice, LPS stimulates more robust production and processing of IL-1β from microglia primed by Amyloid pathology. Astrocytes primed by amyloid pathology also respond in an exaggerated way to exogenous IL-1β, producing enhanced levels of chemokines. If levels of IL-1, arising endogenously from microglial stimulation, are already exaggerated this will produce a further amplification upon stimulating astrocytes that are hypersensitive to IL-1. These exaggerated responses also appear to affect many other astrocytic transcripts involved in innate immune function as shown by fold-change expression in comparison to WT+saline.

Journal: bioRxiv

Article Title: Microglial and Astrocyte priming in the APP/PS1 model of Alzheimer’s Disease: increased vulnerability to acute inflammation and cognitive deficits

doi: 10.1101/344218

Figure Lengend Snippet: Exogenous i.c. administration of LPS in normal mice induces activation of microglial cells to produce regulated secretion of IL-1β and this IL-1β may activate astrocytes to release chemokines such as CCL2, CXCL1 or CXCL10. However, in APP/PS1 mice, LPS stimulates more robust production and processing of IL-1β from microglia primed by Amyloid pathology. Astrocytes primed by amyloid pathology also respond in an exaggerated way to exogenous IL-1β, producing enhanced levels of chemokines. If levels of IL-1, arising endogenously from microglial stimulation, are already exaggerated this will produce a further amplification upon stimulating astrocytes that are hypersensitive to IL-1. These exaggerated responses also appear to affect many other astrocytic transcripts involved in innate immune function as shown by fold-change expression in comparison to WT+saline.

Article Snippet: For confocal microscopy, sections were labelled with the following combinations: 6E10 (1:1000; Biolegend 803001) with an Alexa Fluor 488 anti-mouse (1:800; Invitrogen) together with Iba-1 (1:2000; Abcam ab5076) Alexa Fluor 594 anti-goat (1:800; Invitrogen); 6E10 with an Alexa Fluor 633 anti-mouse (1:800; Invitrogen) in combination with IL-1β (1:50; Peprotech 500-P51) with an Alexa Fluor 488 anti-rabbit (1:800; Invitrogen); 6E10 with an Alexa Fluor 633 anti-mouse and GFAP with an Alexa Fluor 488 anti-rabbit; GFAP (1:2000; Dako Z0334) with an Alexa Fluor 594 anti-rabbit (1:800; Invitrogen) together with IL-1β (1:50; Peprotech 500-P51) with an Alexa Fluor 488 anti-rabbit; GFAP with an Alexa Fluor 488 anti-rabbit in combination with CCL2 (1:200; R&D AF-479-NA) with Alexa Fluor 594 anti-goat; Iba-1 (1:2000; Abcam ab5076) with Alexa Fluor 594 anti-goat.

Techniques: Activation Assay, Amplification, Expressing, Comparison, Saline