monoclonal antibody  (Cedarlane)

Journal: Journal of Lipid Research

Article Title: Aggressive cholesterol lowering normalizes atherosclerosis regression in Jak2 V617F mice

doi: 10.1016/j.jlr.2026.101003

Figure Lengend Snippet: Atherosclerosis regression is incomplete in Jak2 VF MPN mice with moderate cholesterol lowering. A: Study design created with BioRender.com. B: Plasma cholesterol (n = 34, 55, 55, 54, 16, 16 for Ctrl mice, n = 31, 55, 49, 39, 17, 16 for Jak2 VF mice, for weeks 0, 5, 11, 15, 17.5, 21 respectively). P = 0.028, <0.0001, <0.0001, <0.0001, 0.0005, <0.0001 (Ctrl vs. Jak2 VF at weeks 0, 5, 11, 15, 17.5, and 21, respectively). C: H&E images of aortic root lesions. Black lines , necrotic core. Scale bar, 200 μm. D: Lesion area, n = 15–23. P < 0.0001 (Ctrl Baseline vs. Jak2 VF Baseline; Ctrl LDL Lowering vs. Jak2 VF LDL Lowering), P = 0.052 ( Jak2 VF Baseline vs. LDL Lowering). E: Necrotic core area, n = 15–23. P = 0.0079 (Ctrl Baseline vs. Jak2 VF Baseline), P = 0.0002 (Ctrl LDL Lowering vs. Jak2 VF LDL Lowering), P = 0.0003 ( Jak2 VF Baseline vs. LDL Lowering). F: Picrosirius red-stained aortic root lesions. Scale bar, 200 μm. G: Collagen area as a percentage of lesion area, n = 15–24. P = 0.02 (Ctrl Baseline vs. LDL Lowering). H: Images of aortic root lesions stained for MAC2 ( Green ) and DAPI ( Blue ). Scale bar, 200 μm. I: Macrophage area, n = 15–24. P = 0.006 (Ctrl Baseline vs. LDL Lowering), P = 0.16 ( Jak2 VF Baseline vs. LDL Lowering). All quantifications shown as mean ± s.e.m. Two-way ANOVA with the Geisser-Greenhouse correction for sphericity and Tukey’s multiple comparisons test (B). Two-way ANOVA with Tukey’s multiple comparisons test (D, E, G, and I). DAPI, 4′,6-diamidino-2-phenylindole; LDL, low-density lipoprotein; MPN, myeloproliferative neoplasm.

Article Snippet: Following blocking, sections were incubated with the following primary antibodies at the indicated concentrations overnight at 4 °C in a humidified chamber: Absent in melanoma 2 (AIM2) (Abcam, ab119791, 1:250), Cleaved GasD (Cell Signaling, 10137, 10 μg/ml), c-Myc (Cell Signaling, 5605, 5.76 μg/ml), Ki67 (Abcam, ab15580, 9 μg/ml), MAC2 (Cedarlane, CL8942AP 1 μg/ml), MAC2 conjugated to Alexa Fluor 488 (Cedarlane, CL8942AF4, 1 μg/ml), mer proto-oncogene tyrosine kinase (MerTK) (R&D, BAF591, 2 μg/ml), pγH2AX (Cell Signaling, 9718, 0.74 μg/ml), ZsGreen (Thermo Fisher Scientific, TA180002, 10 μg/ml), triggering receptor expressed on myeloid cells 2 (TREM2) (Denali, 4D9 DC1847, 1:100).

Techniques: Clinical Proteomics, Staining

Journal: Journal of Lipid Research

Article Title: Aggressive cholesterol lowering normalizes atherosclerosis regression in Jak2 V617F mice

doi: 10.1016/j.jlr.2026.101003

Figure Lengend Snippet: Aggressive cholesterol lowering normalizes regression in Jak2 VF MPN lesions. A: Study design, created with BioRender.com. B: Plasma cholesterol (n = 5, 38, 19, 18, and 18 for Ctrl mice, n = 5, 37, 15, 16, and 15 for Jak2 VF mice, for weeks 0, 4, 13, 15, and 18, respectively). P = 0.0041 for genotype effect by two-way ANOVA with Geisser-Greenhouse correction, P = 0.011, 0.048, 0.056, and 0.0008 (Ctrl vs. Jak2 VF at weeks 0, 4, 13, and 15, respectively). C: H&E images of aortic root lesions. Black lines , necrotic core. Scale bar, 200 μm. D: Lesion area, n = 13–20. P < 0.0001 (Ctrl Baseline vs. Jak2 VF Baseline; Ctrl LDL Lowering vs. Jak2 VF LDL Lowering). Figures 1 and 4 represent independent regression cohorts performed separately; therefore, absolute lesion area should be compared within each cohort rather than between figures. E: Necrotic core area, n = 13–20. P = 0.0003 (Ctrl Baseline vs. Jak2 VF Baseline), P = 0.0006 (Ctrl LDL Lowering vs. Jak2 VF LDL Lowering), P = 0.90 ( Jak2 VF Baseline vs. LDL Lowering). F: Picrosirius red-stained aortic root lesions. Scale bar, 200 μm. G: Collagen area as a percentage of lesion area, n = 13–20. P < 0.0001 (Ctrl Baseline vs. LDL Lowering), P = 0.0002 ( Jak2 VF Baseline vs. LDL Lowering), P = 0.02 (Ctrl LDL Lowering vs. Jak2 VF LDL Lowering). H: Images of aortic root lesions stained for MAC2 ( Green ) and DAPI ( Blue ). Scale bar, 200 μm. I: Macrophage area, n = 13–20. P < 0.0001 (Ctrl and Jak2 VF Baseline vs. LDL Lowering). All quantifications shown as mean ± s.e.m. Two-way ANOVA with Tukey’s multiple comparisons test (D, G, and I). Two-way ANOVA with the Geisser-Greenhouse correction for sphericity and Tukey’s multiple comparisons test (B). Kruskal-Wallis test with Dunn’s multiple comparisons test (E). DAPI, 4′,6-diamidino-2-phenylindole; LDL, low-density lipoprotein; MPN, myeloproliferative neoplasm.

Article Snippet: Following blocking, sections were incubated with the following primary antibodies at the indicated concentrations overnight at 4 °C in a humidified chamber: Absent in melanoma 2 (AIM2) (Abcam, ab119791, 1:250), Cleaved GasD (Cell Signaling, 10137, 10 μg/ml), c-Myc (Cell Signaling, 5605, 5.76 μg/ml), Ki67 (Abcam, ab15580, 9 μg/ml), MAC2 (Cedarlane, CL8942AP 1 μg/ml), MAC2 conjugated to Alexa Fluor 488 (Cedarlane, CL8942AF4, 1 μg/ml), mer proto-oncogene tyrosine kinase (MerTK) (R&D, BAF591, 2 μg/ml), pγH2AX (Cell Signaling, 9718, 0.74 μg/ml), ZsGreen (Thermo Fisher Scientific, TA180002, 10 μg/ml), triggering receptor expressed on myeloid cells 2 (TREM2) (Denali, 4D9 DC1847, 1:100).

Techniques: Clinical Proteomics, Staining

Journal: Journal of Lipid Research

Article Title: Aggressive cholesterol lowering normalizes atherosclerosis regression in Jak2 V617F mice

doi: 10.1016/j.jlr.2026.101003

Figure Lengend Snippet: Cholesterol lowering suppresses Jak2 VF macrophage proliferation and DNA damage. A: Study design created with BioRender.com. B: Plasma cholesterol (n = 79, 78, and 78 for Baseline mice for weeks 3, 11, and 15, respectively; n = 13, 14 for Ctrl Progression mice, n = 16, 16 for Ctrl LDL Lowering mice, n = 15, 15 for Jak2 VF Progression mice, n = 16, 16 for Jak2 VF LDL Lowering mice, for weeks 18 and 23, respectively). C: Images of aortic root lesions for MAC2 ( Green ), Cleaved GasD ( Red ), and DAPI ( Blue ). Scale bar, 60 μm. D: Log 10 transformed cleaved GasD mean fluorescence intensity (MFI) in lesions with the addition of constant 1, n = 14–16. P = 0.01 (Baseline vs. Jak2 VF Progression), P = 0.051 ( Jak2 VF Progression vs. Jak2 VF LDL Lowering). E: Log 10 transformed cleaved GasD MFI in the necrotic core with the addition of constant 1, n = 14–16. P = 0.04 (Baseline vs. Jak2 VF Progression), P = 0.013 ( Jak2 VF Progression vs. Jak2 VF LDL Lowering). F: Images of aortic root lesions stained for MAC2 ( Green ), Cleaved GasD ( Red ), and DAPI ( Blue ). Scale bar, 60 μm. White arrows , cleaved GasD + macrophages. G: Log 10 transformed cleaved GasD positive macrophages per section with the addition of constant 1, n = 14–16. H: Images of aortic root lesions for MAC2 ( Red ), Ki67 ( White ), and ZsGreen ( Green ). Scale bar, 60 μm. White arrows , macrophages double positive for Ki67 and ZsGreen. I: Macrophages positive for both Ki67 and ZsGreen per section, n = 13–15. P = 0.0019 (Baseline vs. Jak2 VF Progression), P = 0.0006 (Ctrl Progression vs. Jak2 VF Progression), P = 0.0003 ( Jak2 VF Progression vs. Jak2 VF LDL Lowering). J: Macrophages positive for Ki67 but negative for ZsGreen per section, n = 13–16. K: Images of aortic root lesions stained for pγH2AX ( Red ), ZsGreen ( Green ), and DAPI ( Blue ). Scale bar, 60 μm. White arrows, pγH2AX positive cells. L: Log 10 transformed cells double positive for pγH2AX and ZsGreen in lesions with the addition of constant 1, n = 13–16. P = 0.0024 (Baseline vs. Jak2 VF Progression), P = 0.0019 ( Jak2 VF Progression vs. Jak2 VF LDL Lowering). M: Log 10 transformed cells positive for pγH2AX but negative for ZsGreen in lesions with the addition of constant 1, n = 13–16. All quantifications shown as mean ± s.e.m. One-way ANOVA with Holm–Sidak’s multiple comparisons test (D and E). Kruskal–Wallis test with Dunn’s multiple comparison’s test (G, I, J, L, and M). DAPI, 4′,6-diamidino-2-phenylindole; GasD, gasdermin D; LDL, low-density lipoprotein; pγH2AX, phosphorylated histone H2A.X.

Article Snippet: Following blocking, sections were incubated with the following primary antibodies at the indicated concentrations overnight at 4 °C in a humidified chamber: Absent in melanoma 2 (AIM2) (Abcam, ab119791, 1:250), Cleaved GasD (Cell Signaling, 10137, 10 μg/ml), c-Myc (Cell Signaling, 5605, 5.76 μg/ml), Ki67 (Abcam, ab15580, 9 μg/ml), MAC2 (Cedarlane, CL8942AP 1 μg/ml), MAC2 conjugated to Alexa Fluor 488 (Cedarlane, CL8942AF4, 1 μg/ml), mer proto-oncogene tyrosine kinase (MerTK) (R&D, BAF591, 2 μg/ml), pγH2AX (Cell Signaling, 9718, 0.74 μg/ml), ZsGreen (Thermo Fisher Scientific, TA180002, 10 μg/ml), triggering receptor expressed on myeloid cells 2 (TREM2) (Denali, 4D9 DC1847, 1:100).

Techniques: Clinical Proteomics, Transformation Assay, Fluorescence, Staining

Journal: Journal of Lipid Research

Article Title: Aggressive cholesterol lowering normalizes atherosclerosis regression in Jak2 V617F mice

doi: 10.1016/j.jlr.2026.101003

Figure Lengend Snippet: Moderate cholesterol lowering reverses macrophage AIM2 inflammasome activation, DNA damage, and proliferation in Jak2 VF MPN lesions. A: Images of aortic root lesions stained for MAC2 ( Green ), Cleaved GasD ( Red ), and DAPI ( Blue ). Scale bar, 60 μm. B: Log 10 transformed cleaved GasD mean fluorescence intensity (MFI) in lesions with the addition of constant 1, n = 15–24. P = 0.021 (Ctrl Baseline vs. Jak2 VF Baseline). C: Log 10 transformed cleaved GasD mean fluorescence intensity (MFI) in necrotic cores with the addition of constant 1, n = 15–24. P = 0.0008 (Ctrl Baseline vs. Jak2 VF Baseline), P = 0.032 (Ctrl LDL Lowering vs. Jak2 VF LDL Lowering). D: Images of aortic root lesions stained for MAC2 ( Green ), Cleaved GasD ( Red ), and DAPI ( Blue ). Scale bar, 60 μm. White arrows , cleaved GasD + macrophages. E: Log 10 transformed cleaved GasD positive macrophages per section with the addition of constant 1, n = 15–24. P = 0.024 ( Jak2 VF Baseline vs. LDL Lowering). F: Images of aortic root lesions stained for MAC2 ( Green ), AIM2 ( Red ), and DAPI ( Blue ). Scale bar, 60 μm. White arrows , AIM2 positive macrophages. G: Log 10 transformed AIM2 positive macrophages per section with the addition of constant 1, n = 15–23. P = 0.0001 (Ctrl Baseline vs. Jak2 VF Baseline), P = 0.03 ( Jak2 VF Baseline vs. LDL Lowering). H: Images of aortic root lesions stained for MAC2 ( Green ), pγH2AX ( Red ), and DAPI ( Blue ). Scale bar, 60 μm. White arrows , pγH2AX positive cells. I: Log 10 transformed pγH2AX positive cells per section with the addition of constant 1, n = 15–23. P = 0.0019 (Ctrl Baseline vs. Jak2 VF Baseline), P = 0.045 ( Jak2 VF Baseline vs. LDL Lowering). J: Images of aortic root lesions stained for MAC2 ( Green ), Ki67 ( Red ), and DAPI ( Blue ). Scale bar, 60 μm. White arrows , Ki67 positive macrophages. K: Log 10 transformed Ki67 positive macrophages per section with the addition of constant 1, n = 15–23. P = 0.0019 ( Jak2 VF Baseline vs. LDL Lowering). MAC2 intensity differences reflect independent staining and imaging sessions across panels. All quantifications shown as mean ± s.e.m. Two-way ANOVA with Tukey’s multiple comparisons test (B and C). Kruskal–Wallis test with Dunn’s multiple comparisons test (E, G, I, and K). AIM2, absent in melanoma 2; DAPI, 4′,6-diamidino-2-phenylindole; GasD, gasdermin D; LDL, low-density lipoprotein; MPN, myeloproliferative neoplasm; pγH2AX, phosphorylated histone H2A.X.

Article Snippet: Following blocking, sections were incubated with the following primary antibodies at the indicated concentrations overnight at 4 °C in a humidified chamber: Absent in melanoma 2 (AIM2) (Abcam, ab119791, 1:250), Cleaved GasD (Cell Signaling, 10137, 10 μg/ml), c-Myc (Cell Signaling, 5605, 5.76 μg/ml), Ki67 (Abcam, ab15580, 9 μg/ml), MAC2 (Cedarlane, CL8942AP 1 μg/ml), MAC2 conjugated to Alexa Fluor 488 (Cedarlane, CL8942AF4, 1 μg/ml), mer proto-oncogene tyrosine kinase (MerTK) (R&D, BAF591, 2 μg/ml), pγH2AX (Cell Signaling, 9718, 0.74 μg/ml), ZsGreen (Thermo Fisher Scientific, TA180002, 10 μg/ml), triggering receptor expressed on myeloid cells 2 (TREM2) (Denali, 4D9 DC1847, 1:100).

Techniques: Activation Assay, Staining, Transformation Assay, Fluorescence, Imaging

Journal: Journal of Lipid Research

Article Title: Aggressive cholesterol lowering normalizes atherosclerosis regression in Jak2 V617F mice

doi: 10.1016/j.jlr.2026.101003

Figure Lengend Snippet: Moderate cholesterol lowering reverses impaired efferocytosis in Jak2 VF MPN lesions while increasing TREM2 Hi macrophages in control and Jak2 VF MPN lesions. A: Images of aortic root lesions stained for MAC2 ( Green ), MerTK ( Red ), and DAPI ( Blue ). Scale bar, 60 μm. B: Percentage of MAC2 positive area double positive for MerTK and MAC2, n = 15–23. P = 0.02 (Ctrl Baseline vs. Jak2 VF Baseline), P < 0.0001 ( Jak2 VF Baseline vs. LDL Lowering). C: Images of aortic root lesions stained for MAC2 ( Green ), TREM2 ( Red ), and DAPI ( Blue ). Scale bar, 60 μm. D: Percentage of MAC2 positive area double positive for TREM2 and MAC2, n = 15–24. P < 0.0001 (Ctrl and Jak2 VF Baseline vs. LDL Lowering). E: Images of in situ efferocytosis in aortic root lesions: MAC2 ( Green ), TUNEL ( Red ), and DAPI ( Blue ). Scale bar, 60 μm. White arrows , nuclei double positive for TUNEL and MAC2. White wedges , nuclei positive for TUNEL but negative for MAC2. F: Percentage of TUNEL positive nuclei also positive for MAC2. P = 0.011 (Ctrl Baseline vs. Jak2 VF Baseline), P = 0.021 ( Jak2 VF Baseline vs. LDL Lowering). MAC2 intensity differences reflect independent staining and imaging sessions across panels, n = 13–24. All quantifications shown as mean ± s.e.m. Two-way ANOVA with Tukey’s multiple comparisons test (D). Kruskal-Wallis test with Dunn’s multiple comparisons test (B and F). DAPI, 4′,6-diamidino-2-phenylindole; LDL, low-density lipoprotein; MerTK, mer proto-oncogene tyrosine kinase; MPN, myeloproliferative neoplasm; TREM2, triggering receptor expressed on myeloid cells 2.

Article Snippet: Following blocking, sections were incubated with the following primary antibodies at the indicated concentrations overnight at 4 °C in a humidified chamber: Absent in melanoma 2 (AIM2) (Abcam, ab119791, 1:250), Cleaved GasD (Cell Signaling, 10137, 10 μg/ml), c-Myc (Cell Signaling, 5605, 5.76 μg/ml), Ki67 (Abcam, ab15580, 9 μg/ml), MAC2 (Cedarlane, CL8942AP 1 μg/ml), MAC2 conjugated to Alexa Fluor 488 (Cedarlane, CL8942AF4, 1 μg/ml), mer proto-oncogene tyrosine kinase (MerTK) (R&D, BAF591, 2 μg/ml), pγH2AX (Cell Signaling, 9718, 0.74 μg/ml), ZsGreen (Thermo Fisher Scientific, TA180002, 10 μg/ml), triggering receptor expressed on myeloid cells 2 (TREM2) (Denali, 4D9 DC1847, 1:100).

Techniques: Control, Staining, In Situ, TUNEL Assay, Imaging

Journal: Journal of Lipid Research

Article Title: Aggressive cholesterol lowering normalizes atherosclerosis regression in Jak2 V617F mice

doi: 10.1016/j.jlr.2026.101003

Figure Lengend Snippet: Aggressive cholesterol lowering more strongly decreases macrophage pyroptosis and DNA damage in Jak2 VF MPN lesions. A: Images of aortic root lesions stained for MAC2 ( Green ), cleaved GasD ( Red ), and DAPI ( Blue ). Scale bar, 60 μm. B: Log 10 transformed cleaved GasD MFI in the necrotic core with the addition of constant 1, n = 14–20. P = 0.0064 for genotype effect and P = 0.021 for treatment effect by two-way ANOVA. C: Images of aortic root lesions stained for MAC2 ( Green ), Cleaved GasD ( Red ), and DAPI ( Blue ). Scale bar, 60 μm. White arrows , cleaved GasD + macrophages. D: Log 10 transformed cleaved GasD positive macrophages per section with the addition of constant 1, n = 13–20. P = 0.023 (Ctrl Baseline vs. Jak2 VF Baseline), P = 0.005 ( Jak2 VF Baseline vs. LDL Lowering). E: Representative immunoblot analysis of full-length and cleaved GasD in CD11b + splenocytes. F: Log 10 transformed densitometric quantification of the ratio of cleaved GasD to full-length GasD from E with the addition of constant 1. n = 7–8 biological replicates. P = 0.0003 (Ctrl Baseline vs. Jak2 VF Baseline), P = 0.0046 ( Jak2 VF Baseline vs. LDL Lowering). G: Images of aortic root lesions stained for MAC2 ( Green ), pγH2AX ( Red ), and DAPI ( Blue ). Scale bar, 60 μm. White arrows , pγH2AX positive cells. H: Log 10 transformed pγH2AX positive cells per section with the addition of constant 1, n = 14–20. P = 0.016 (Ctrl Baseline vs. LDL Lowering), P = 0.0016 (Ctrl Baseline vs. Jak2 VF Baseline), P < 0.0001 ( Jak2 VF Baseline vs. LDL Lowering). All quantifications shown as mean ± s.e.m. Two-way ANOVA with Tukey’s multiple comparisons test (B, D, F, and H). DAPI, 4′,6-diamidino-2-phenylindole; GasD, gasdermin D; LDL, low-density lipoprotein; MFI, mean fluorescence intensity; MPN, myeloproliferative neoplasm; pγH2AX, phosphorylated histone H2A.X.

Article Snippet: Following blocking, sections were incubated with the following primary antibodies at the indicated concentrations overnight at 4 °C in a humidified chamber: Absent in melanoma 2 (AIM2) (Abcam, ab119791, 1:250), Cleaved GasD (Cell Signaling, 10137, 10 μg/ml), c-Myc (Cell Signaling, 5605, 5.76 μg/ml), Ki67 (Abcam, ab15580, 9 μg/ml), MAC2 (Cedarlane, CL8942AP 1 μg/ml), MAC2 conjugated to Alexa Fluor 488 (Cedarlane, CL8942AF4, 1 μg/ml), mer proto-oncogene tyrosine kinase (MerTK) (R&D, BAF591, 2 μg/ml), pγH2AX (Cell Signaling, 9718, 0.74 μg/ml), ZsGreen (Thermo Fisher Scientific, TA180002, 10 μg/ml), triggering receptor expressed on myeloid cells 2 (TREM2) (Denali, 4D9 DC1847, 1:100).

Techniques: Staining, Transformation Assay, Western Blot, Fluorescence

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