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ikaros inhibitor lenalidomide  (MedChemExpress)


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    MedChemExpress ikaros inhibitor lenalidomide
    A , B SerpinB2 was quantified in THP-1 macrophages after stimulation with IFN-γ in the presence or absence of Bay-118072, an NF-kB inhibitor, by qPCR ( A ) (n = 8/group) and confocal imaging ( B ) (n = 10/group). A , B each dot represents cells cultured in one well. C VAT from lean and obese mice were stained for the markers of fibroblasts, macrophages, and adipocytes along with IFN-γ. The frequencies of IFN-γ-expressing cells were assessed by confocal microscopy (n = 5/group) Scale bar=20 µm. D – J LysM +/+ Ifngr1 fl/fl and LysM cre/+ Ifngr1 fl/fl mice were fed with an HFD for four months. D VAT resident macrophages, SerpinB2-expressing macrophages, and caspase 3 + resident macrophages were measured by confocal microscopy (n = 12/group). E The frequencies of apoptotic (annexin V + PI − ) macrophages were measured by flow cytometry (n = 5 for WT and 4 for KO). F GTT and ITT were performed, and fasting blood glucose, serum insulin, lipid concentrations, and body weights were evaluated (n = 10/group, combined data of at least 2 independent experiments). G , H Immunoblot showing pAkt, total Akt, Glut4, adiponectin, and Ppar y expressions in muscle, visceral adipose. and liver (n = 6/group). I , J qPCR was carried out to measure the expression of the metabolic and inflammatory genes in VAT (n = 7/group). K , L The top four transcription factors predicted to bind to the SerpinB2 promoter ( K ), and ChIP validation of <t>Ikaros</t> binding to the SerpinB2 promoter ( L ) (n = 4/group, with two sets of primers) are shown. M Ikaros quantification by qPCR in BMDM cultured in the presence or absence of 250 milli units IFN-γ (n = 10 and 8 for with and without IFN-γ, respectively). L , M each dot represents cells cultured in one well. N SerpinB2 expression in BMDM treated with or without IFN-γ and <t>lenalidomide</t> (representative image of the 3 independent experiments). O Ifngr1 and Ifngr2 were quantified by RNA seq in the adipose macrophage subsets (n = 3/group, each dot represents one mouse.). P – R Evaluation of apoptosis in BMDM lacking SerpinB2 (P) (n = 6/group), THP-1 macrophages overexpressing SerpinB2 ( Q ) (n = 4/group), and VAT resident macrophages in obese SerpinB2 +/+ and SerpinB2 − / − mice ( R ) (n = 4 for WT and 6 for KO) by flow cytometry. * P < 0.05, ** P < 0.01, *** P < 0.001. The Mann Whitney test (two-tailed) was used to determine the significance between two groups. One-way ANOVA with Bonferoni’s post hoc correction test was performed to determine differences among data obtained from more than two groups (Fig. 4B).
    Ikaros Inhibitor Lenalidomide, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 30 article reviews
    ikaros inhibitor lenalidomide - by Bioz Stars, 2026-09
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    Images

    1) Product Images from "Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation"

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation

    Journal: Nature Communications

    doi: 10.1038/s41467-026-69196-4

    A , B SerpinB2 was quantified in THP-1 macrophages after stimulation with IFN-γ in the presence or absence of Bay-118072, an NF-kB inhibitor, by qPCR ( A ) (n = 8/group) and confocal imaging ( B ) (n = 10/group). A , B each dot represents cells cultured in one well. C VAT from lean and obese mice were stained for the markers of fibroblasts, macrophages, and adipocytes along with IFN-γ. The frequencies of IFN-γ-expressing cells were assessed by confocal microscopy (n = 5/group) Scale bar=20 µm. D – J LysM +/+ Ifngr1 fl/fl and LysM cre/+ Ifngr1 fl/fl mice were fed with an HFD for four months. D VAT resident macrophages, SerpinB2-expressing macrophages, and caspase 3 + resident macrophages were measured by confocal microscopy (n = 12/group). E The frequencies of apoptotic (annexin V + PI − ) macrophages were measured by flow cytometry (n = 5 for WT and 4 for KO). F GTT and ITT were performed, and fasting blood glucose, serum insulin, lipid concentrations, and body weights were evaluated (n = 10/group, combined data of at least 2 independent experiments). G , H Immunoblot showing pAkt, total Akt, Glut4, adiponectin, and Ppar y expressions in muscle, visceral adipose. and liver (n = 6/group). I , J qPCR was carried out to measure the expression of the metabolic and inflammatory genes in VAT (n = 7/group). K , L The top four transcription factors predicted to bind to the SerpinB2 promoter ( K ), and ChIP validation of Ikaros binding to the SerpinB2 promoter ( L ) (n = 4/group, with two sets of primers) are shown. M Ikaros quantification by qPCR in BMDM cultured in the presence or absence of 250 milli units IFN-γ (n = 10 and 8 for with and without IFN-γ, respectively). L , M each dot represents cells cultured in one well. N SerpinB2 expression in BMDM treated with or without IFN-γ and lenalidomide (representative image of the 3 independent experiments). O Ifngr1 and Ifngr2 were quantified by RNA seq in the adipose macrophage subsets (n = 3/group, each dot represents one mouse.). P – R Evaluation of apoptosis in BMDM lacking SerpinB2 (P) (n = 6/group), THP-1 macrophages overexpressing SerpinB2 ( Q ) (n = 4/group), and VAT resident macrophages in obese SerpinB2 +/+ and SerpinB2 − / − mice ( R ) (n = 4 for WT and 6 for KO) by flow cytometry. * P < 0.05, ** P < 0.01, *** P < 0.001. The Mann Whitney test (two-tailed) was used to determine the significance between two groups. One-way ANOVA with Bonferoni’s post hoc correction test was performed to determine differences among data obtained from more than two groups (Fig. 4B).
    Figure Legend Snippet: A , B SerpinB2 was quantified in THP-1 macrophages after stimulation with IFN-γ in the presence or absence of Bay-118072, an NF-kB inhibitor, by qPCR ( A ) (n = 8/group) and confocal imaging ( B ) (n = 10/group). A , B each dot represents cells cultured in one well. C VAT from lean and obese mice were stained for the markers of fibroblasts, macrophages, and adipocytes along with IFN-γ. The frequencies of IFN-γ-expressing cells were assessed by confocal microscopy (n = 5/group) Scale bar=20 µm. D – J LysM +/+ Ifngr1 fl/fl and LysM cre/+ Ifngr1 fl/fl mice were fed with an HFD for four months. D VAT resident macrophages, SerpinB2-expressing macrophages, and caspase 3 + resident macrophages were measured by confocal microscopy (n = 12/group). E The frequencies of apoptotic (annexin V + PI − ) macrophages were measured by flow cytometry (n = 5 for WT and 4 for KO). F GTT and ITT were performed, and fasting blood glucose, serum insulin, lipid concentrations, and body weights were evaluated (n = 10/group, combined data of at least 2 independent experiments). G , H Immunoblot showing pAkt, total Akt, Glut4, adiponectin, and Ppar y expressions in muscle, visceral adipose. and liver (n = 6/group). I , J qPCR was carried out to measure the expression of the metabolic and inflammatory genes in VAT (n = 7/group). K , L The top four transcription factors predicted to bind to the SerpinB2 promoter ( K ), and ChIP validation of Ikaros binding to the SerpinB2 promoter ( L ) (n = 4/group, with two sets of primers) are shown. M Ikaros quantification by qPCR in BMDM cultured in the presence or absence of 250 milli units IFN-γ (n = 10 and 8 for with and without IFN-γ, respectively). L , M each dot represents cells cultured in one well. N SerpinB2 expression in BMDM treated with or without IFN-γ and lenalidomide (representative image of the 3 independent experiments). O Ifngr1 and Ifngr2 were quantified by RNA seq in the adipose macrophage subsets (n = 3/group, each dot represents one mouse.). P – R Evaluation of apoptosis in BMDM lacking SerpinB2 (P) (n = 6/group), THP-1 macrophages overexpressing SerpinB2 ( Q ) (n = 4/group), and VAT resident macrophages in obese SerpinB2 +/+ and SerpinB2 − / − mice ( R ) (n = 4 for WT and 6 for KO) by flow cytometry. * P < 0.05, ** P < 0.01, *** P < 0.001. The Mann Whitney test (two-tailed) was used to determine the significance between two groups. One-way ANOVA with Bonferoni’s post hoc correction test was performed to determine differences among data obtained from more than two groups (Fig. 4B).

    Techniques Used: Imaging, Cell Culture, Staining, Expressing, Confocal Microscopy, Flow Cytometry, Western Blot, Biomarker Discovery, Binding Assay, RNA Sequencing, MANN-WHITNEY, Two Tailed Test

    Related Articles

    Imaging:

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMs were treated with or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN- γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation.
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMswere treatedwith or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN-γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.

    Cell Culture:

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMs were treated with or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN- γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation.
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMswere treatedwith or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN-γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.

    Staining:

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMs were treated with or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN- γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation.
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMswere treatedwith or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN-γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.

    Expressing:

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMs were treated with or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN- γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation.
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMswere treatedwith or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN-γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.

    Confocal Microscopy:

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMs were treated with or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN- γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation.
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMswere treatedwith or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN-γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.

    Flow Cytometry:

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMs were treated with or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN- γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation.
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMswere treatedwith or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN-γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.

    Western Blot:

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMs were treated with or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN- γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation.
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMswere treatedwith or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN-γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.

    Biomarker Discovery:

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMs were treated with or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN- γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation.
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMswere treatedwith or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN-γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.

    Binding Assay:

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMs were treated with or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN- γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation.
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMswere treatedwith or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN-γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.

    RNA Sequencing:

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMs were treated with or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN- γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation.
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMswere treatedwith or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN-γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.

    MANN-WHITNEY:

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMs were treated with or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN- γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation.
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMswere treatedwith or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN-γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.

    Two Tailed Test:

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN- γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250 μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMs were treated with or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN- γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 10 5 /ml and differentiated into macrophages in the presence of 100 nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucose media in 6-well plates or 4-well chambered slides as described previously , .

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation.
    Article Snippet: Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.Similarly, BMDM from bone marrow cells of C57BL/6 mice were cultured and treated with 250 milliunits of IFN-γ (Invitrogen, #PHC4031) or various chemicals such as palmitate (Acros Organics, #416700050), cycloheximide (Alfa Aesar, #J6690103), and 250μM mitotempol (Fisher Scientific, #NC1229394) for 0-24 hours in either 6 well plates or chamber slides.. In a separate experiment, BMDMswere treatedwith or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN-γ for 24 hours.. THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.THP-1 cell culture: THP-1 monocytes were seeded at a density of 2 × 105/ml and differentiated into macrophages in the presence of 100nM Phorbol-12-myristate-13-acetate (PMA) (Calbiochem, # 524400) for 2 days in RPMI low glucosemedia in 6-well plates or 4-well chambered slides as described previously44,165.



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    ikaros inhibitor lenalidomide  (MedChemExpress)
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    MedChemExpress ikaros inhibitor lenalidomide
    A , B SerpinB2 was quantified in THP-1 macrophages after stimulation with IFN-γ in the presence or absence of Bay-118072, an NF-kB inhibitor, by qPCR ( A ) (n = 8/group) and confocal imaging ( B ) (n = 10/group). A , B each dot represents cells cultured in one well. C VAT from lean and obese mice were stained for the markers of fibroblasts, macrophages, and adipocytes along with IFN-γ. The frequencies of IFN-γ-expressing cells were assessed by confocal microscopy (n = 5/group) Scale bar=20 µm. D – J LysM +/+ Ifngr1 fl/fl and LysM cre/+ Ifngr1 fl/fl mice were fed with an HFD for four months. D VAT resident macrophages, SerpinB2-expressing macrophages, and caspase 3 + resident macrophages were measured by confocal microscopy (n = 12/group). E The frequencies of apoptotic (annexin V + PI − ) macrophages were measured by flow cytometry (n = 5 for WT and 4 for KO). F GTT and ITT were performed, and fasting blood glucose, serum insulin, lipid concentrations, and body weights were evaluated (n = 10/group, combined data of at least 2 independent experiments). G , H Immunoblot showing pAkt, total Akt, Glut4, adiponectin, and Ppar y expressions in muscle, visceral adipose. and liver (n = 6/group). I , J qPCR was carried out to measure the expression of the metabolic and inflammatory genes in VAT (n = 7/group). K , L The top four transcription factors predicted to bind to the SerpinB2 promoter ( K ), and ChIP validation of <t>Ikaros</t> binding to the SerpinB2 promoter ( L ) (n = 4/group, with two sets of primers) are shown. M Ikaros quantification by qPCR in BMDM cultured in the presence or absence of 250 milli units IFN-γ (n = 10 and 8 for with and without IFN-γ, respectively). L , M each dot represents cells cultured in one well. N SerpinB2 expression in BMDM treated with or without IFN-γ and <t>lenalidomide</t> (representative image of the 3 independent experiments). O Ifngr1 and Ifngr2 were quantified by RNA seq in the adipose macrophage subsets (n = 3/group, each dot represents one mouse.). P – R Evaluation of apoptosis in BMDM lacking SerpinB2 (P) (n = 6/group), THP-1 macrophages overexpressing SerpinB2 ( Q ) (n = 4/group), and VAT resident macrophages in obese SerpinB2 +/+ and SerpinB2 − / − mice ( R ) (n = 4 for WT and 6 for KO) by flow cytometry. * P < 0.05, ** P < 0.01, *** P < 0.001. The Mann Whitney test (two-tailed) was used to determine the significance between two groups. One-way ANOVA with Bonferoni’s post hoc correction test was performed to determine differences among data obtained from more than two groups (Fig. 4B).
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    A , B SerpinB2 was quantified in THP-1 macrophages after stimulation with IFN-γ in the presence or absence of Bay-118072, an NF-kB inhibitor, by qPCR ( A ) (n = 8/group) and confocal imaging ( B ) (n = 10/group). A , B each dot represents cells cultured in one well. C VAT from lean and obese mice were stained for the markers of fibroblasts, macrophages, and adipocytes along with IFN-γ. The frequencies of IFN-γ-expressing cells were assessed by confocal microscopy (n = 5/group) Scale bar=20 µm. D – J LysM +/+ Ifngr1 fl/fl and LysM cre/+ Ifngr1 fl/fl mice were fed with an HFD for four months. D VAT resident macrophages, SerpinB2-expressing macrophages, and caspase 3 + resident macrophages were measured by confocal microscopy (n = 12/group). E The frequencies of apoptotic (annexin V + PI − ) macrophages were measured by flow cytometry (n = 5 for WT and 4 for KO). F GTT and ITT were performed, and fasting blood glucose, serum insulin, lipid concentrations, and body weights were evaluated (n = 10/group, combined data of at least 2 independent experiments). G , H Immunoblot showing pAkt, total Akt, Glut4, adiponectin, and Ppar y expressions in muscle, visceral adipose. and liver (n = 6/group). I , J qPCR was carried out to measure the expression of the metabolic and inflammatory genes in VAT (n = 7/group). K , L The top four transcription factors predicted to bind to the SerpinB2 promoter ( K ), and ChIP validation of Ikaros binding to the SerpinB2 promoter ( L ) (n = 4/group, with two sets of primers) are shown. M Ikaros quantification by qPCR in BMDM cultured in the presence or absence of 250 milli units IFN-γ (n = 10 and 8 for with and without IFN-γ, respectively). L , M each dot represents cells cultured in one well. N SerpinB2 expression in BMDM treated with or without IFN-γ and lenalidomide (representative image of the 3 independent experiments). O Ifngr1 and Ifngr2 were quantified by RNA seq in the adipose macrophage subsets (n = 3/group, each dot represents one mouse.). P – R Evaluation of apoptosis in BMDM lacking SerpinB2 (P) (n = 6/group), THP-1 macrophages overexpressing SerpinB2 ( Q ) (n = 4/group), and VAT resident macrophages in obese SerpinB2 +/+ and SerpinB2 − / − mice ( R ) (n = 4 for WT and 6 for KO) by flow cytometry. * P < 0.05, ** P < 0.01, *** P < 0.001. The Mann Whitney test (two-tailed) was used to determine the significance between two groups. One-way ANOVA with Bonferoni’s post hoc correction test was performed to determine differences among data obtained from more than two groups (Fig. 4B).

    Journal: Nature Communications

    Article Title: Tissue-resident macrophage survival depends on mitochondrial function regulated by SerpinB2 in chronic inflammation

    doi: 10.1038/s41467-026-69196-4

    Figure Lengend Snippet: A , B SerpinB2 was quantified in THP-1 macrophages after stimulation with IFN-γ in the presence or absence of Bay-118072, an NF-kB inhibitor, by qPCR ( A ) (n = 8/group) and confocal imaging ( B ) (n = 10/group). A , B each dot represents cells cultured in one well. C VAT from lean and obese mice were stained for the markers of fibroblasts, macrophages, and adipocytes along with IFN-γ. The frequencies of IFN-γ-expressing cells were assessed by confocal microscopy (n = 5/group) Scale bar=20 µm. D – J LysM +/+ Ifngr1 fl/fl and LysM cre/+ Ifngr1 fl/fl mice were fed with an HFD for four months. D VAT resident macrophages, SerpinB2-expressing macrophages, and caspase 3 + resident macrophages were measured by confocal microscopy (n = 12/group). E The frequencies of apoptotic (annexin V + PI − ) macrophages were measured by flow cytometry (n = 5 for WT and 4 for KO). F GTT and ITT were performed, and fasting blood glucose, serum insulin, lipid concentrations, and body weights were evaluated (n = 10/group, combined data of at least 2 independent experiments). G , H Immunoblot showing pAkt, total Akt, Glut4, adiponectin, and Ppar y expressions in muscle, visceral adipose. and liver (n = 6/group). I , J qPCR was carried out to measure the expression of the metabolic and inflammatory genes in VAT (n = 7/group). K , L The top four transcription factors predicted to bind to the SerpinB2 promoter ( K ), and ChIP validation of Ikaros binding to the SerpinB2 promoter ( L ) (n = 4/group, with two sets of primers) are shown. M Ikaros quantification by qPCR in BMDM cultured in the presence or absence of 250 milli units IFN-γ (n = 10 and 8 for with and without IFN-γ, respectively). L , M each dot represents cells cultured in one well. N SerpinB2 expression in BMDM treated with or without IFN-γ and lenalidomide (representative image of the 3 independent experiments). O Ifngr1 and Ifngr2 were quantified by RNA seq in the adipose macrophage subsets (n = 3/group, each dot represents one mouse.). P – R Evaluation of apoptosis in BMDM lacking SerpinB2 (P) (n = 6/group), THP-1 macrophages overexpressing SerpinB2 ( Q ) (n = 4/group), and VAT resident macrophages in obese SerpinB2 +/+ and SerpinB2 − / − mice ( R ) (n = 4 for WT and 6 for KO) by flow cytometry. * P < 0.05, ** P < 0.01, *** P < 0.001. The Mann Whitney test (two-tailed) was used to determine the significance between two groups. One-way ANOVA with Bonferoni’s post hoc correction test was performed to determine differences among data obtained from more than two groups (Fig. 4B).

    Article Snippet: In a separate experiment, BMDMs were treated with or without Ikaros inhibitor lenalidomide (5 μM) (MedChemExpress, #HYA0003) for 2 hours followed by stimulation with 250 milliunits of IFN- γ for 24 hours.

    Techniques: Imaging, Cell Culture, Staining, Expressing, Confocal Microscopy, Flow Cytometry, Western Blot, Biomarker Discovery, Binding Assay, RNA Sequencing, MANN-WHITNEY, Two Tailed Test