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l-arginine free dmem medium cat # cc927-0500  (GeneDireX Inc)
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GeneDireX Inc l-arginine free dmem medium cat # cc927-0500
ATG5 deficiency in eosinophils induces Arg2 expression, resulting in decreased beiging in adipocytes. (A, B) RNA-seq. The volcano plot of differentially expressed genes (DEGs) in bone marrow (A) and blood (B) -isolated eosinophils from Atg5 eoΔ Il5 tg was compared to bone marrow and blood-isolated eosinophils from Ctrl Il5 tg . A total of 13 and 25 DEGs were identified, respectively. (C) Quantitative PCR. The isolation of eosinophils from bone marrow (BM) and blood of Ctrl II5 tg and Atg5 eoΔ II5 tg mice was followed by gene expression analysis for Arg2 and Atg5 . The expression of the target genes was normalized using Actb and Nono as reference genes (n=3). (D) Immunoblotting. Protein lysates from freshly isolated BM and blood-derived eosinophils of Ctrl II5 tg and Atg5 eoΔ II5 tg mice were subjected to analysis for ATG5 and ARG2 protein expression. ACTB protein levels were utilized as loading controls. (E) Arginase activity assay. Eosinophils freshly isolated from BM and blood samples of Ctrl II5 tg and Atg5 eoΔ II5 tg mice were subjected to an arginase activity assay (n = 3). (F) Immunoblotting. Protein lysates were obtained from mature adipocytes incubated in L-arginine free <t>DMEM</t> supplemented with different L-arginine concentrations for five hours. ACTB protein levels served as loading controls. (G) Immunoblotting. Eosinophils were cultured in the presence and absence of ABH-hydrochloride, and their supernatants were applied to adipocytes, followed by UCP1 immunoblotting. (H) Quantitative PCR. qPCR was performed on adipocytes that underwent the same treatments described in the previous section to evaluate the expression level of thermogenic genes. Expression levels were normalized using Actb and Nono as reference genes and compared to control mice. (n = 3). Values are means ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001.
L Arginine Free Dmem Medium Cat # Cc927 0500, supplied by GeneDireX Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ATG5 deficiency in eosinophils induces Arg2 expression, resulting in decreased beiging in adipocytes. (A, B) RNA-seq. The volcano plot of differentially expressed genes (DEGs) in bone marrow (A) and blood (B) -isolated eosinophils from Atg5 eoΔ Il5 tg was compared to bone marrow and blood-isolated eosinophils from Ctrl Il5 tg . A total of 13 and 25 DEGs were identified, respectively. (C) Quantitative PCR. The isolation of eosinophils from bone marrow (BM) and blood of Ctrl II5 tg and Atg5 eoΔ II5 tg mice was followed by gene expression analysis for Arg2 and Atg5 . The expression of the target genes was normalized using Actb and Nono as reference genes (n=3). (D) Immunoblotting. Protein lysates from freshly isolated BM and blood-derived eosinophils of Ctrl II5 tg and Atg5 eoΔ II5 tg mice were subjected to analysis for ATG5 and ARG2 protein expression. ACTB protein levels were utilized as loading controls. (E) Arginase activity assay. Eosinophils freshly isolated from BM and blood samples of Ctrl II5 tg and Atg5 eoΔ II5 tg mice were subjected to an arginase activity assay (n = 3). (F) Immunoblotting. Protein lysates were obtained from mature adipocytes incubated in L-arginine free DMEM supplemented with different L-arginine concentrations for five hours. ACTB protein levels served as loading controls. (G) Immunoblotting. Eosinophils were cultured in the presence and absence of ABH-hydrochloride, and their supernatants were applied to adipocytes, followed by UCP1 immunoblotting. (H) Quantitative PCR. qPCR was performed on adipocytes that underwent the same treatments described in the previous section to evaluate the expression level of thermogenic genes. Expression levels were normalized using Actb and Nono as reference genes and compared to control mice. (n = 3). Values are means ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001.

Journal: Frontiers in Immunology

Article Title: The regulatory role of eosinophils in adipose tissue depends on autophagy

doi: 10.3389/fimmu.2023.1331151

Figure Lengend Snippet: ATG5 deficiency in eosinophils induces Arg2 expression, resulting in decreased beiging in adipocytes. (A, B) RNA-seq. The volcano plot of differentially expressed genes (DEGs) in bone marrow (A) and blood (B) -isolated eosinophils from Atg5 eoΔ Il5 tg was compared to bone marrow and blood-isolated eosinophils from Ctrl Il5 tg . A total of 13 and 25 DEGs were identified, respectively. (C) Quantitative PCR. The isolation of eosinophils from bone marrow (BM) and blood of Ctrl II5 tg and Atg5 eoΔ II5 tg mice was followed by gene expression analysis for Arg2 and Atg5 . The expression of the target genes was normalized using Actb and Nono as reference genes (n=3). (D) Immunoblotting. Protein lysates from freshly isolated BM and blood-derived eosinophils of Ctrl II5 tg and Atg5 eoΔ II5 tg mice were subjected to analysis for ATG5 and ARG2 protein expression. ACTB protein levels were utilized as loading controls. (E) Arginase activity assay. Eosinophils freshly isolated from BM and blood samples of Ctrl II5 tg and Atg5 eoΔ II5 tg mice were subjected to an arginase activity assay (n = 3). (F) Immunoblotting. Protein lysates were obtained from mature adipocytes incubated in L-arginine free DMEM supplemented with different L-arginine concentrations for five hours. ACTB protein levels served as loading controls. (G) Immunoblotting. Eosinophils were cultured in the presence and absence of ABH-hydrochloride, and their supernatants were applied to adipocytes, followed by UCP1 immunoblotting. (H) Quantitative PCR. qPCR was performed on adipocytes that underwent the same treatments described in the previous section to evaluate the expression level of thermogenic genes. Expression levels were normalized using Actb and Nono as reference genes and compared to control mice. (n = 3). Values are means ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: To assess the effect of different concentrations of L-arginine, mature adipocytes incubated in L-arginine free DMEM medium (Cat # CC927-0500, GeneDireX, Keelung, Taiwan) supplemented with various concentrations of L-arginine (Cat # A6969, Sigma-Aldrich) for five hours.

Techniques: Expressing, RNA Sequencing, Isolation, Real-time Polymerase Chain Reaction, Gene Expression, Western Blot, Derivative Assay, Arginase Activity Assay, Incubation, Cell Culture, Control