rat anti gal3 mac2  (Cedarlane)

Journal: Neuroprotection

Article Title: Environmental enrichment modulates chronic poststroke inflammation and links white matter TREM2‐positive microglia in recovery in mice

doi: 10.1002/nep3.70028

Figure Lengend Snippet: Infarct size correlations with peri‐infarct immunofluorescence analysis of ionized calcium binding adaptor molecule 1 (Iba1), galectin‐3 (Gal3), and purinergic receptor P2Y12 (P2RY12). (A) Representative immunostaining of 4′,6‐diamidino‐2‐phenylindole (DAPI), Iba1, P2RY12, and Gal3 in a standard environment (SE) mouse at peri‐infarct area (at ×20 magnification). (B) Representative immunostaining of DAPI, Iba1, P2RY12, and Gal3 in an enriched environment (EE) mouse at peri‐infarct area (at ×20 magnification). (C) Quantification of indirect infarct area measurements. (D) Correlation of infarct area with Neuroscore per group. (E) Iba1 coverage quantification measured as the percentage of image covered by Iba1 area (%area). (F) Correlation of infarct area with Iba1 coverage. (G) Gal3 coverage quantification measured as the percentage of image covered by Iba1 + Gal3 + area (%area). (H) Correlation of infarct area with Gal3 coverage. (I) P2RY12 coverage quantification measured as the percentage of image covered by Iba1 + P2RY12 + area (%area). (J) Correlation of infarct area with P2RY12 coverage. Peri‐infarct area is shown as dashed red lines in A and B. In (C, E, G, and I), values are expressed as individual experimental replicates with mean ± SEM. In (D, F, H, and J), values are expressed as individual experimental replicates with simple linear regression lines; Pearson correlation's r value with p ‐value are also shown. In (C, E, G, and I), unpaired t ‐test was performed. n = 4/6 mice in SE and n = 7 mice in EE (2 mice in SE were not behaviorally characterized). P ‐values and r values are expressed with 3 decimals. P ‐values were not corrected for multiple comparisons.

Article Snippet: They were then incubated at 4°C overnight with one of the following antibodies: Iba1 (1:500, rabbit, Cat# 016‐26721; Wako), Gal3 (1:750, goat, Cat# AF1197; R&D Systems) P2RY12 (1:200, rat, Cat# S16007D; Biolegend).

Techniques: Immunofluorescence, Binding Assay, Immunostaining

Journal: Neuroprotection

Article Title: Environmental enrichment modulates chronic poststroke inflammation and links white matter TREM2‐positive microglia in recovery in mice

doi: 10.1002/nep3.70028

Figure Lengend Snippet: Infarct size correlations with white matter immunofluorescence analysis of ionized calcium binding adaptor molecule 1 (Iba1), galectin‐3 (Gal3), and purinergic receptor P2Y12 (P2RY12). (A) Representative immunostaining of 4′,6‐diamidino‐2‐phenylindole (DAPI), Iba1, P2RY12, and Gal3 (at ×20 magnification) in a standard environment (SE) mouse at white matter area (corpus callosum + external capsule). (B) Representative immunostaining of DAPI, Iba1, P2RY12, and Gal3 in an enriched environment (EE) mouse at white matter area (at ×20 magnification). (C) Iba1 coverage quantification measured as the percentage of image covered by Iba1 area (%area). (D) Gal3 coverage quantification measured as the percentage of image covered by Iba1 + Gal3 + area (%area). (E) P2RY12 coverage quantification measured as the percentage of image covered by Iba1 + P2RY12 + area (%area). (F) Correlation of infarct area with Iba1 coverage. (G) Correlation of infarct area with Gal3 coverage. (H) Correlation of infarct area with P2RY12 coverage. White matter area is shown as dashed red lines in (A, B). In (C–E) values are expressed as individual experimental replicates with mean ± SEM. In (F–H) values are expressed as individual experimental replicates with simple linear regression lines; Pearson correlation's r value with p ‐value are also shown. In (C–E) unpaired t ‐test was performed n = 6 mice in SE and n = 7 mice in EE. p ‐Values and r values are expressed with 3 decimals. p ‐Values were not corrected for multiple comparisons.

Article Snippet: They were then incubated at 4°C overnight with one of the following antibodies: Iba1 (1:500, rabbit, Cat# 016‐26721; Wako), Gal3 (1:750, goat, Cat# AF1197; R&D Systems) P2RY12 (1:200, rat, Cat# S16007D; Biolegend).

Techniques: Immunofluorescence, Binding Assay, Immunostaining

Journal: Neuroprotection

Article Title: Environmental enrichment modulates chronic poststroke inflammation and links white matter TREM2‐positive microglia in recovery in mice

doi: 10.1002/nep3.70028

Figure Lengend Snippet: Quantification of peri‐infarct myelin debris, white matter myelin loss, and their correlations with microglial markers. (A) Representative myelin staining in a standard environment mouse (at ×20 magnification). (B) Enlarged views of infarct contralateral cortical (green square) and peri‐infarct (red square) myelin in a standard environment (SE) mouse. (C) Representative myelin staining in an enriched environment mouse (at ×20 magnification). (D) Enlarged views of infarct contralateral cortical (green square) and peri‐infarct (red square) myelin in an enriched environment (EE) mouse. (E) Myelin debris coverage quantification measured as the percentage of peri‐infarct image covered by Black Gold Myelin dark debris area (%area). (F) Correlation of infarct area with myelin debris coverage. (G) Myelin loss quantification measured as the percentage of myelin lost at corpus callosum in ipsilateral versus contralateral infarct. (H) Correlation of infarct area with myelin loss. (I) Correlations of myelin debris with ionized calcium binding adaptor molecule 1 (Iba1), galectin‐3 (Gal3), purinergic receptor P2Y12 (P2RY12), cluster of differentiation 68 (CD68), and triggering receptor expressed on myeloid cells 2 (TREM2) coverages at peri‐infarct. (J) Correlations of myelin loss with Iba1, Gal3, P2RY12, CD68, and TREM2 coverages at white matter. In (E, G) values are expressed as individual experimental replicates with mean ± SEM. In (F, H, I, J), values are expressed as individual experimental replicates with simple linear regression lines; Pearson correlation's r value with p ‐value are also shown. In (E, G) unpaired t‐test was performed. n = 6 mice in SE and n = 7 mice in EE. p ‐Values and r values are expressed with 3 decimals. p ‐Values were not corrected for multiple comparisons.

Article Snippet: They were then incubated at 4°C overnight with one of the following antibodies: Iba1 (1:500, rabbit, Cat# 016‐26721; Wako), Gal3 (1:750, goat, Cat# AF1197; R&D Systems) P2RY12 (1:200, rat, Cat# S16007D; Biolegend).

Techniques: Staining, Binding Assay

Journal: In vivo (Athens, Greece)

Article Title: Expression of Galectin 3 and Activating Transcription Factor 3 in Nigral Dopaminergic Neurons of 6-Hydroxydopamine Induced Parkinsonian Rat Model.

doi: 10.21873/invivo.13938

Figure Lengend Snippet: Figure 2. Gal3 expression was upregulated in the ipsilateral nigral dopaminergic neurons. (A) Photographs of Gal3 immunohistochemistry in the contralateral (Cont) and ipsilateral (Ispi) substantia nigra (SN) after 6‐hydroxydopamine (6‐OHDA) lesion. Gal3 immunoreactivity was upregulated in the ipsilateral SN. (B) High magnification image of the area designated by the black square inside of (A). Morphologically typical multipolar neurons with large cell body and many prominent processes (arrows) and neuroglia with small cell bodies and many long and slender radiating processes (arrowheads) were positive for Gal3 in the ipsilateral SN. (C) Double immunofluorescence images of Gal3 and TH and their merged image, in the ipsilateral SN at 5 dpl. Gal3 was expressed in many nigral dopaminergic neurons (arrows). (D) Double immunofluorescence images of Gal3 and Iba1 and their merged image. Gal3 was also expressed in many microglial cells (arrows). (E) Double immunofluorescence images of Gal3 and GFAP and their merged image. None of the cells immunolabeled for Gal3 were colocalized with GFAP‐positive astrocytes (arrows). (F) Quantitative analysis showing the average number of neurons expressing Gal3 per unit area (mm2) at each time point. Scale bars represent 200 μm in (A), 50 μm in (B and E). Data are expressed as means±SEM (n=6, *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001). dpl, Days post‐lesion.

Article Snippet: Antibody Catalog Manufacturer Host Description Concentration number IHC IF Gal3 AF‐1197 R&D system Goat Wide range of cell type 1:500 1:50 ATF3 Sc‐188 Santa Cruz Rabbit Wide range of cell type 1:1000 1:100 TH MAB5280 Millipore Mouse Tyrosine hydroxylase 1:1000 1:500 Iba1 AB283319 Abcam Mouse Microglia marker 1:200 GFAP AB279289 Abcam Mouse Astrocyte marker 1:200 FG AB153 Millipore Rabbit Fluorogold 1:500 1:100 Gal3, Galectin 3; ATF3, activating transcription factor 3; TH, tyrosine hydroxylase; Iba1, ionized calcium‐binding adapter molecule 1; GFAP, glial fibrillary acidic protein; FG, fluorogold; IHC, immunohistochemistry; IF, immunofluorescence. antibodies against TH (1:500), or rabbit polyclonal antibodies against FG (1:100).

Techniques: Expressing, Immunohistochemistry, Immunofluorescence, Immunolabeling

Journal: In vivo (Athens, Greece)

Article Title: Expression of Galectin 3 and Activating Transcription Factor 3 in Nigral Dopaminergic Neurons of 6-Hydroxydopamine Induced Parkinsonian Rat Model.

doi: 10.21873/invivo.13938

Figure Lengend Snippet: Figure 4. Co‐localization of Gal3 and ATF3 in the 6‐OHDA insulted dopaminergic neurons. (A) Representative photomicrographs showing triple immunofluorescence labeling for FG, Gal3, and ATF3 and their merged image in the ipsilateral SN after 6‐OHDA lesion at 5 dpl. (B) High magnification image of the area designated by the white square inside of (A). ATF3 and Gal3 were colocalized in the dopaminergic neurons that are labeled with FG, i.e., neurons that have been retrogradely insulted with 6‐OHDA. Scale bar represents 200 μm and 50 μm in (A) and (B), respectively. dpl, Days post‐lesion; Ipsi, ipsilateral.

Article Snippet: Antibody Catalog Manufacturer Host Description Concentration number IHC IF Gal3 AF‐1197 R&D system Goat Wide range of cell type 1:500 1:50 ATF3 Sc‐188 Santa Cruz Rabbit Wide range of cell type 1:1000 1:100 TH MAB5280 Millipore Mouse Tyrosine hydroxylase 1:1000 1:500 Iba1 AB283319 Abcam Mouse Microglia marker 1:200 GFAP AB279289 Abcam Mouse Astrocyte marker 1:200 FG AB153 Millipore Rabbit Fluorogold 1:500 1:100 Gal3, Galectin 3; ATF3, activating transcription factor 3; TH, tyrosine hydroxylase; Iba1, ionized calcium‐binding adapter molecule 1; GFAP, glial fibrillary acidic protein; FG, fluorogold; IHC, immunohistochemistry; IF, immunofluorescence. antibodies against TH (1:500), or rabbit polyclonal antibodies against FG (1:100).

Techniques: Immunofluorescence, Labeling