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mac2  (Cedarlane)


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    Structured Review

    Cedarlane mac2
    Mac2, supplied by Cedarlane, used in various techniques. Bioz Stars score: 96/100, based on 398 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+mac2/Anti-Mouse%2FHuman+Mac-2+(Galectin-3)%2C+Purified+(Clone+M3%2F38)+(rat+IgG2a)/pm41794821-312-23-25
    Average 96 stars, based on 398 article reviews
    mac2 - by Bioz Stars, 2026-09
    96/100 stars

    Images

    Related Articles

    Fluorescence:

    Article Title: Human resistin is critical to activation of the NLRP3 inflammasome in macrophages
    Article Snippet: .. After deparaffinization of tissue, rehydration, and antigen retrieval, sections were treated with anti-NLRP3 (Abcam, ab214185) and anti-Mac2 (Cedarlane, CL8942LE), anti-MPO (R&D Systems, AF3667), anti-CD79b (Abcam, ab134147), anti-BTK (Sigma-Aldrich, SAB4502936), anti-hResistin (R&D Systems, AF1359), or anti-RELMα (R&D Systems, MAB1523) antibodies overnight at 4°C and then with Alexa Fluor 488-donkey anti-mouse IgG (Jackson ImmunoResearch, 715-545-150) and Cy3-donkey anti-rabbit IgG (Jackson ImmunoResearch, 711-166-152) for double fluorescence staining. .. Sections were then incubated with the appropriate fluorochrome-coupled secondary antibody (Jackson ImmunoResearch).

    Staining:

    Article Title: Human resistin is critical to activation of the NLRP3 inflammasome in macrophages
    Article Snippet: .. After deparaffinization of tissue, rehydration, and antigen retrieval, sections were treated with anti-NLRP3 (Abcam, ab214185) and anti-Mac2 (Cedarlane, CL8942LE), anti-MPO (R&D Systems, AF3667), anti-CD79b (Abcam, ab134147), anti-BTK (Sigma-Aldrich, SAB4502936), anti-hResistin (R&D Systems, AF1359), or anti-RELMα (R&D Systems, MAB1523) antibodies overnight at 4°C and then with Alexa Fluor 488-donkey anti-mouse IgG (Jackson ImmunoResearch, 715-545-150) and Cy3-donkey anti-rabbit IgG (Jackson ImmunoResearch, 711-166-152) for double fluorescence staining. .. Sections were then incubated with the appropriate fluorochrome-coupled secondary antibody (Jackson ImmunoResearch).

    Article Title: Sex-specific angiogenic responses in endothelial cells—role of the pluripotency factor OCT4
    Article Snippet: .. To assess macrophage infiltration within injured skin, immunofluorescence staining was performed using an anti-MAC2 [Cedarlane; CL8942AP; 1:500] antibody with 4,6-diamidino-2-phenylindole (DAPI). ..

    Article Title: Lipid lowering alone fails to limit atherosclerosis progression and neutrophilic inflammation in middle-aged mice
    Article Snippet: In addition, for Mac2 and αSMA stains, sections were permeabilized in a solution containing 0.01%-0.05% Triton X-100 (cat# 9036-19-5, Sigma Aldrich Inc, St. Louis, MO, USA) prior to blocking. .. After brief washes, samples were stained with either anti-Mac2 (cat# CL8942AP, Cedarlane Labs, Burlington, ON, USA) at 1:10000, anti-Histone H3 (citrulline R2 + R8 + R17) (cat#AB281584, clone RM1001, Abcam, Waltham, MA, USA) at 1:500, or anti-smooth muscle actin (cat#. 23081-1-AP, Proteintech, Rosemont, IL, USA) at 1:1000 in 1%BSA. .. Secondary antibodies used were Alexa Fluor 594 anti-rat (cat# A-11007, Thermo Fisher Scientific, Carlsbad, CA, USA) (Mac2) and Alexa Fluor 594 anti-rabbit (cat# A-11037, Thermo Fisher Scientific, Carlsbad, CA, USA) (H3Cit and αSMA).

    Article Title: Chronic Glaucoma Induced in Rats by a Single Injection of Fibronectin-Loaded PLGA Microspheres: IOP-Dependent and IOP-Independent Neurodegeneration.
    Article Snippet: .. Sections were stained with haematoxylin/eosin and mounted with DPX mounting medium or incubated overnight at 4 ◦C with the following primary antibodies: anti-fibronectin (AB2033, Chemicon), 1:100; anti-Brn3a (14A6, Santa Cruz Biotechnology, Dallas, TX, USA), 1:50; anti-GFAP (Z0334, Agilent, Dako, Santa Clara, CA, USA), 1:500; anti-Mac2 (CL8942LE, Cedarlane, Burlington, Canada) 1:50. ..

    Incubation:

    Article Title: Loss of TIM4-Dependent Efferocytosis in Kupffer Cells Promotes Liver Fibrosis in Nonalcoholic Steatohepatitis
    Article Snippet: For immunofluorescence experiments, human liver sections were incubated at 4 °C overnight with anti-human TIM4 (Cell signaling technology, #75484T, RRID:AB_2799871, 1:200 dilution), anti-CD68 (Agilent, #GA60961-2, RRID:AB_2661840, 1:500 dilution), and anti-cleaved caspase3 (Cell signaling technology, # 9661, RRID:AB_2341188, 1:100 dilution) in PBS containing 1% donkey serum. .. Mouse liver sections were incubated at 4 °C overnight with anti-mouse TIM4 (RD, #AF2826, RRID:AB_2201844, 1:200 dilution), anti-Clec4f (RD, #MAB2784, RRID:AB_2081338, 1:250 dilution), anti-Mac2 (Cedarlane, #CL8942AP, RRID:AB_10060357, 1:500 dilution), anti-F4/80 (Cell signaling technology, #70076s, RRID:AB_2799771, 1:200), anti-osteopontin (Opn, RD, #AF808, RRID:AB_2194992, 1:100 dilution), anti-α-smooth muscle actin (Sigma, #C6198, RRID:AB_476856, 1:100 dilution) and anti-cleaved caspase3 (Cell signaling technology, # 9661, RRID:AB_2341188, 1:100 dilution) in PBS containing 1% donkey serum. ..

    Article Title: Nucleoporin93 limits Yap activity to prevent endothelial cell senescence
    Article Snippet: .. Adjacent tissue sections were incubated with the following primary antibodies (4°C, overnight): anti‐Nup93 (1:500, Lusk Lab, Yale University); anti‐γH2AX (1:200, Sigma‐Aldrich, 05‐636); anti‐Mac2 (1:100, Cedarlane, CL8942AP); anti‐Yap (1:200, Santa Cruz, sc101199); anti‐VWF (1:200, Abcam, ab11713). ..

    Article Title: Chronic Glaucoma Induced in Rats by a Single Injection of Fibronectin-Loaded PLGA Microspheres: IOP-Dependent and IOP-Independent Neurodegeneration.
    Article Snippet: .. Sections were stained with haematoxylin/eosin and mounted with DPX mounting medium or incubated overnight at 4 ◦C with the following primary antibodies: anti-fibronectin (AB2033, Chemicon), 1:100; anti-Brn3a (14A6, Santa Cruz Biotechnology, Dallas, TX, USA), 1:50; anti-GFAP (Z0334, Agilent, Dako, Santa Clara, CA, USA), 1:500; anti-Mac2 (CL8942LE, Cedarlane, Burlington, Canada) 1:50. ..

    Immunofluorescence:

    Article Title: Sex-specific angiogenic responses in endothelial cells—role of the pluripotency factor OCT4
    Article Snippet: .. To assess macrophage infiltration within injured skin, immunofluorescence staining was performed using an anti-MAC2 [Cedarlane; CL8942AP; 1:500] antibody with 4,6-diamidino-2-phenylindole (DAPI). ..

    Immunohistochemistry:

    Article Title: The context-dependent epigenetic and organogenesis programs determine 3D vs. 2D cellular fitness of MYC-driven murine liver cancer cells
    Article Snippet: Antibody , Anti-ACTIN (Mouse, monoclonal) , Sigma , Cat# A1978, RRID: AB_4766 , WB (1:5000). .. Antibody , Anti-MAC2 (Rat, monoclonal) , Accurate/CEDARLANE , Cat# CL-8942AP, RRID: AB_10060357 , IHC (1:1000). .. Recombinant DNA reagent , LeveV2-vector , Addgene , 52961 , .



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    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 <t>MAC2</t> ( 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.
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    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 <t>MAC2</t> ( 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.
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    Cedarlane anti mac2
    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 <t>MAC2</t> ( 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.
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    Podocyte damage and inflammatory changes are worsened by HPD in systemic GC-A knockout mice. ( a ) Immunohistochemical (IHC) staining for WT-1 in glomerulus. The graph shows the number of WT-1-positive cells per glomerulus as quantitative analysis. ( b ) IF staining for nephrin in each group. Scale Bar represents 50 μm. ( c ) Representative images of electron microscopic analysis in each group. The graphs indicate quantitative analysis of foot process width and GBM thickness. Scale Bar represents 1 μm. ( d ) IHC staining of <t>MAC2</t> in glomerulus (▶). The graph shows the number of MAC2-positive macrophages per glomerulus. Scale Bar represents 50 μm. ( e , f ) Glomerular mRNA expression levels of Tgfb1 , Ctgf , Serpine1 , Ccl2 , Emr1 , Icam1 , and Vcam1 in each group. ( g ) Renal mRNA expression levels of Agt , Ren1 , and Sgk1 in each group. β-Actin ( Actb ) was used as internal control. Values are expressed as means ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, NS; not significant.
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    Image Search Results


    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    Podocyte damage and inflammatory changes are worsened by HPD in systemic GC-A knockout mice. ( a ) Immunohistochemical (IHC) staining for WT-1 in glomerulus. The graph shows the number of WT-1-positive cells per glomerulus as quantitative analysis. ( b ) IF staining for nephrin in each group. Scale Bar represents 50 μm. ( c ) Representative images of electron microscopic analysis in each group. The graphs indicate quantitative analysis of foot process width and GBM thickness. Scale Bar represents 1 μm. ( d ) IHC staining of MAC2 in glomerulus (▶). The graph shows the number of MAC2-positive macrophages per glomerulus. Scale Bar represents 50 μm. ( e , f ) Glomerular mRNA expression levels of Tgfb1 , Ctgf , Serpine1 , Ccl2 , Emr1 , Icam1 , and Vcam1 in each group. ( g ) Renal mRNA expression levels of Agt , Ren1 , and Sgk1 in each group. β-Actin ( Actb ) was used as internal control. Values are expressed as means ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, NS; not significant.

    Journal: Scientific Reports

    Article Title: Finerenone ameliorates diabetic kidney disease exacerbated by deletion of natriuretic peptide/guanylyl cyclase-A signaling and dietary high-protein load

    doi: 10.1038/s41598-025-25362-0

    Figure Lengend Snippet: Podocyte damage and inflammatory changes are worsened by HPD in systemic GC-A knockout mice. ( a ) Immunohistochemical (IHC) staining for WT-1 in glomerulus. The graph shows the number of WT-1-positive cells per glomerulus as quantitative analysis. ( b ) IF staining for nephrin in each group. Scale Bar represents 50 μm. ( c ) Representative images of electron microscopic analysis in each group. The graphs indicate quantitative analysis of foot process width and GBM thickness. Scale Bar represents 1 μm. ( d ) IHC staining of MAC2 in glomerulus (▶). The graph shows the number of MAC2-positive macrophages per glomerulus. Scale Bar represents 50 μm. ( e , f ) Glomerular mRNA expression levels of Tgfb1 , Ctgf , Serpine1 , Ccl2 , Emr1 , Icam1 , and Vcam1 in each group. ( g ) Renal mRNA expression levels of Agt , Ren1 , and Sgk1 in each group. β-Actin ( Actb ) was used as internal control. Values are expressed as means ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, NS; not significant.

    Article Snippet: Primary antibodies used for immunohistochemical and immunofluorescence studies were anti-GC-A antibody (GTX109810, GeneTex, Inc., Irvine, CA, USA), anti-MAC2 antibody (CL8942F, Cedarlane, Ontario, Canada), anti-Wilms tumor 1 (WT1) antibody (sc-15421, Santa Cruz Biotechnology, Dallas, TX), anti-nephrin antibody (AF3159, R&D Systems, Inc., Minneapolis, MN, USA), and anti-Tie2 antibody (AF762, R&D Systems, Inc.).

    Techniques: Knock-Out, Immunohistochemical staining, Immunohistochemistry, Staining, Expressing, Control

    Podocyte damage and profibrotic changes are worsened by HPD in endothelial cell-specific GC-A knockout mice. ( a ) IHC staining of WT-1 in glomerulus. The graph shows the number of WT-1-positive cells per glomerulus as quantitative analysis. Scale Bar represents 50 μm. ( b ) IF staining for nephrin. Scale Bar represents 50 μm. ( c ) Representative images of electron microscopic analysis in each group. The graphs indicate quantitative analysis of foot process width and GBM thickness. Scale Bar represents 1 μm. ( d ) IHC staining of MAC2 in glomerulus (▶). The graph shows the number of MAC2-positive macrophages per glomerulus. Scale Bar represents 100 μm. ( e ) Glomerular mRNA expression levels of Tgfb1 , Ctgf , Fn1 , Serpine1 , Ccl2 , and Emr1 in each group. ( f ) Renal mRNA levels of Fn1 , Serpine1 , Lcn2 , Agt, and Sgk1 in each group. β-Actin ( Actb ) was used as internal control. Values are expressed as means ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, NS; not significant.

    Journal: Scientific Reports

    Article Title: Finerenone ameliorates diabetic kidney disease exacerbated by deletion of natriuretic peptide/guanylyl cyclase-A signaling and dietary high-protein load

    doi: 10.1038/s41598-025-25362-0

    Figure Lengend Snippet: Podocyte damage and profibrotic changes are worsened by HPD in endothelial cell-specific GC-A knockout mice. ( a ) IHC staining of WT-1 in glomerulus. The graph shows the number of WT-1-positive cells per glomerulus as quantitative analysis. Scale Bar represents 50 μm. ( b ) IF staining for nephrin. Scale Bar represents 50 μm. ( c ) Representative images of electron microscopic analysis in each group. The graphs indicate quantitative analysis of foot process width and GBM thickness. Scale Bar represents 1 μm. ( d ) IHC staining of MAC2 in glomerulus (▶). The graph shows the number of MAC2-positive macrophages per glomerulus. Scale Bar represents 100 μm. ( e ) Glomerular mRNA expression levels of Tgfb1 , Ctgf , Fn1 , Serpine1 , Ccl2 , and Emr1 in each group. ( f ) Renal mRNA levels of Fn1 , Serpine1 , Lcn2 , Agt, and Sgk1 in each group. β-Actin ( Actb ) was used as internal control. Values are expressed as means ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, NS; not significant.

    Article Snippet: Primary antibodies used for immunohistochemical and immunofluorescence studies were anti-GC-A antibody (GTX109810, GeneTex, Inc., Irvine, CA, USA), anti-MAC2 antibody (CL8942F, Cedarlane, Ontario, Canada), anti-Wilms tumor 1 (WT1) antibody (sc-15421, Santa Cruz Biotechnology, Dallas, TX), anti-nephrin antibody (AF3159, R&D Systems, Inc., Minneapolis, MN, USA), and anti-Tie2 antibody (AF762, R&D Systems, Inc.).

    Techniques: Knock-Out, Immunohistochemistry, Staining, Expressing, Control