mac2 (also known as galectin-3)  (Galectin Therapeutics)

Journal: bioRxiv

Article Title: A Mac2-positive progenitor-like microglial population survives independent of CSF1R signaling in adult mouse brain

doi: 10.1101/722090

Figure Lengend Snippet: ( a ) Volcano plot showing differentially expressed genes identified in cluster-4. Lgals3 , which encodes Mac2 protein is highlighted. Upregulated DEGs are colored in red and downregulated DEGs are colored in blue. ( b ) tSNE plot showing expression distribution of Lgals3 in all clusters. Log-transformed total UMI was plotted. ( c ) Violin plot showing expression of Lgals3 in all clusters. ( d ) Representative confocal images showing colocalization of Iba1 and Mac2 in native mice (Ctrl) and PLX-treated mice (D0, 2 weeks of PLX diet). Images were collected from the hippocampal region (shown in mini-map). Solid box highlights cell that is Iba1+Mac2+ and dotted box highlights cell that is Iba1+Mac2-. Enlarged images from the boxed area are shown in separate channels (panel i-v). ( e ) Quantification of Iba1+Mac2-cell numbers. Unpaired t test was used. ( f ) Quantification of Iba1+Mac2+ cell numbers. Mann Whitney test was used. ( g ) Quantification of the percentage of Mac2+ cells among all Iba1+ microglia. Unpaired t test was used. Number of C57/BL6J mice (2.5 – 4 Mo) used (panel e-g): Ctrl (n=5); D0 (n=6). Quantification in (panel e-g) was performed on images (1131.56 μm x 1938.59 μm) collected at the hippocampus using the VERSA automated slide scanner (Leica, 20x lens).

Article Snippet: Mac2, also known as Galectin-3, is a galactoside binding protein highly expressed in myeloid cells that can be secreted to modulate a wide variety of immune functions ( ).

Techniques: Expressing, Transformation Assay, MANN-WHITNEY

Journal: bioRxiv

Article Title: A Mac2-positive progenitor-like microglial population survives independent of CSF1R signaling in adult mouse brain

doi: 10.1101/722090

Figure Lengend Snippet: ( a ) Experimental design of the lineage mapping. Cx3Cr1-CreERT2/STOP flox -RFP mice were injected with tamoxifen (10 days) to label microglia with RFP. Mice are either treated with PLX diet for 3 weeks (PLX 1X ) or underwent repopulation for 2 weeks and treated with PLX diet again for another 3 weeks (PLX 2X ). ( b ) Quantification area was performed on the entire parenchymal region. ( c ) Representative confocal images showing colocalization of Mac2 and RFP expression. Boxed area is enlarged and separated by each channel. Images were collected from the hippocampal region. ( d ) Quantification of the percentage of Mac2+ cell that are RFP+. Number of CX3CR1-CreERT2/STOP flox -RFP mice (7-9 Mo) used: Ctrl (n=3); PLX 1X (n=3); PLX 2X (n=3). One-way ANOVA was used. P-value summary is shown as ns (p > 0.05); * (p ≤ 0.05); ** (p ≤ 0.01); *** (p ≤ 0.001); **** (p ≤ 0.0001). ( e ) Experimental design of microglial repopulation timeline and EdU injections. C57/BL6J mice were treated with PLX diet for 2 weeks (D0) and switched to control diet (CD) to start repopulation for 4 days (D4) or 14 days (D14). EdU was injected on repopulation day 3. ( f ) Brain region used for quantification. Quantification in panel (h-k) was performed on images (1292.23 μm x 1130.7 μm) collected at the Ectorhinal cortex (ECT) using the VERSA automated slide scanner (Leica, 20x lens). ( g ) Representative confocal images showing immunofluorescence staining of EdU (yellow), Iba1 (cyan), and Mac2 (magenta) in the ectorhinal region. Boxed area is shown by separated channels at the bottom. ( h ) Quantification of Iba1+Mac2-cells in the ectorhinal cortex. ( i ) Quantification of Iba1+Mac2+ cells in the ectorhinal cortex. ( j ) Quantification of the percentage of Iba1+ microglia that are Mac2+ in the ectorhinal cortex. ( k ) Quantification of the percentage of EdU+ labeling in either Iba1+Mac2-cells (blue bar) or Iba1+Mac2+ cells (red bar). Number of C57/BL6 mice (2-3.5 Mo) used: Ctrl (n=5); D0 (n=4); D4 (n=5); D14 (n=5). Statistical tests used: 1) In panels (h-j), one-way ANOVA with Dunnett’s multiple comparisons test was used to compare with Ctrl; 2) In panels (k), two-way ANOVA with Dunnett’s multiple comparisons test was used to compare with Ctrl for each cell population. P-value summary is shown as ns (p > 0.05); * (p ≤ 0.05); ** (p ≤ 0.01); *** (p ≤ 0.001); **** (p ≤ 0.0001).

Article Snippet: Mac2, also known as Galectin-3, is a galactoside binding protein highly expressed in myeloid cells that can be secreted to modulate a wide variety of immune functions ( ).

Techniques: Injection, Expressing, Control, Immunofluorescence, Staining, Labeling

Journal: bioRxiv

Article Title: A Mac2-positive progenitor-like microglial population survives independent of CSF1R signaling in adult mouse brain

doi: 10.1101/722090

Figure Lengend Snippet: ( a ) Ridge plot showing isolation of Mac2+ cells from all clusters. Mac2+ cells were separated based on high Lgals3 expression (mean plus one SD, Log10[UMI+1] = 2.172241, shown as vertical dotted line). ( b ) tSNE plot showing the spatial distribution of Mac2+ cells in different clusters. ( c ) Volcano plot showing differentially expressed genes (DEGs) in Mac2+ cells compared to homeostatic microglia (Cluster-1). Upregulated DEGs are colored in red and downregulated DEGs are colored in blue. Genes of interest are highlighted in text. ( d ) Donut chart showing the percentage of microglial developmental genes in all upregulated DEGs in Mac2+ cells (385 genes). ( e ) Donut chart showing the percentage of microglial developmental genes in all downregulated DEGs in Mac2+ cells (443 genes). ( f ) Bar plot showing top-10 hallmark pathways enriched in upregulated Mac2+ DEGs from Gene Set Enrichment Analysis (GSEA). The fraction in the bar shows the number of genes found in the DEGs (numerator) and the number of total genes curated for the corresponding pathway (denominator). ( g ) Same analysis as in (f) but shows enrichment of pathways in downregulated DEGs. ( h ) Dot plot showing enrichment of Wikipathway terms found in upregulated DEGs in Mac2+ cells. The fraction next to the dot shows the number of genes found in the DEGs (numerator) and the number of total genes curated in the corresponding pathway (denominator). ( i ) Same analysis as in (h) but shows enrichment of pathways found in downregulated DEGs.

Article Snippet: Mac2, also known as Galectin-3, is a galactoside binding protein highly expressed in myeloid cells that can be secreted to modulate a wide variety of immune functions ( ).

Techniques: Isolation, Expressing

Journal: bioRxiv

Article Title: A Mac2-positive progenitor-like microglial population survives independent of CSF1R signaling in adult mouse brain

doi: 10.1101/722090

Figure Lengend Snippet: ( a ) Bar graph showing the relative frequency of cells from each treatment group distributed in all Mac2+ cells. A total 45 Mac2+cells were from homeostatic ctrl brain (3.01%). ( b ) Volcano plot showing differentially expressed genes (DEGs) in Mac2+ ctrl cells in comparison to homeostatic microglia (cluster-1). Upregulated DEGs are colored in red while downregulated DEGs are colored in blue. Genes of interest are highlighted in text. ( c ) Venn diagram showing the common upregulated DEGs found between all Mac2+ cells (left circle) and Mac2+ cells from ctrl brain (right circle). ( d ) Venn diagram showing the common downregulated DEGs found between all Mac2+ cells (left circle) and Mac2+ cells from ctrl brain (right circle). ( e ) Donut chart showing the percentage of microglial developmental genes among the upregulated DEGs in Mac2+ cells from ctrl brain (335 genes). ( f ) Donut chart showing the percentage of microglial developmental genes among the downregulated DEGs in Mac2+ cells from ctrl brain (370 genes).

Article Snippet: Mac2, also known as Galectin-3, is a galactoside binding protein highly expressed in myeloid cells that can be secreted to modulate a wide variety of immune functions ( ).

Techniques: Comparison

Journal: PLoS Genetics

Article Title: Cell-Autonomous Progeroid Changes in Conditional Mouse Models for Repair Endonuclease XPG Deficiency

doi: 10.1371/journal.pgen.1004686

Figure Lengend Snippet: (A) Average body weight of C57Bl6/FVB F1 hybrid wild type ( wt ) females (black circles) and forebrain-specific XPG-deficient ( Emx1-Xpg ) females (gray circles); n≥4 animals/group. (B) Onset of clasping of the hind limbs in Emx1-Xpg mice; n = 7 animals/group. (C) Representative images of GFAP immunostained sagittal neocortex sections of 26- and 52-week old Emx1-Xpg and wt mice showing progressive astrocytosis in Emx1-Xpg mice. (D) Representative images of Mac2 immunostained sagittal brain sections of 26- and 52-week old Emx1-Xpg and wt mice showing Mac2-positive microgliosis and a progressive decrease in size of the cerebral cortex and hippocampus of Emx1-Xpg mice. Arrows indicate microgliosis in corpus callosum and fimbria fornix. A thionin counterstaining was used. (E) Quantification of p53-positive cells in neocortex and cerebellum of 26- and 52-week old Emx1-Xpg and wt mice. Values are the average of four sections per genotype. Arrows indicate p53 positive cells. (F) Representative confocal images showing double labeled p53-NeuN cells in the neocortex (left) and p53-S100ß in the fimbria fornix (right) of 26-week old Emx1-Xpg mice. Arrows indicate p53 positive cells. NCx: neocortex, cc: corpus callosum, Str: striatum, ff: fimbria fornix, Hip: hippocampus. Scale bars: 50 µm (C), 500 µm (D), 200 µm (E) and 20 µm (F). Error bars indicate standard error of the mean. **p<0.01.

Article Snippet: Staining for Mac2 (also known as galectin-3), to outline phagocytosing microglia cells , revealed very high levels of Mac2-positive cells in the corpus callosum and the fimbria fornix of Emx1 - Xpg mice ( and ), and a moderate amount of phagocytosing microglia in the neocortex and hippocampus ( ).

Techniques: Labeling