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ccaat enhancer binding protein beta  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc ccaat enhancer binding protein beta
    Ccaat Enhancer Binding Protein Beta, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 151 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/ccaat+enhancer+binding+protein+beta/C%2FEBPbeta+Antibody/pm37331165-69-42-49
    Average 95 stars, based on 151 article reviews
    ccaat enhancer binding protein beta - by Bioz Stars, 2026-09
    95/100 stars

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    Related Articles

    Membrane:

    Article Title: Gene regulation of RMR-related DNAJC6 on adipogenesis and mitochondria function in 3T3-L1 preadipocytes.
    Article Snippet: .. Reactions with the following primary antibodies (diluted 1:500e1:2000) were performed overnight on the membrane at 4 C: DNAJC6 (Cat #PA5-26981; Invitrogen, Waltham,MA, USA), Pref-1 (Cat #sc-376,755; Santa Cruz, Dallas, TX, USA), peroxisome proliferator-activated receptor gamma (PPARg; Cat #2435; CST, Danvers, MA, USA), CCAAT/enhancer binding protein beta (C/EBPb; Cat #3082; CST, Danvers, MA, USA), CCAAT/ enhancer binding protein alpha (C/EBPa; Cat #8178; CST, Danvers, MA, USA), adipocyte protein 2 (aP2; Cat #sc-271,529; Santa Cruz, Dallas, TX, USA), phosphohormone-sensitive lipase (pHSL; Cat #4139; CST, Danvers, MA, USA), hormone-sensitive lipase (HSL; Cat #4107; CST, Danvers, MA, USA), leptin receptor (LEPR; Cat #ab5593; Abcam), leptin (Cat #675002; Biolegend, San Diego, CA, USA), adiponectin (Cat #2789; CST, Danvers, MA, USA), insulin receptor b (Cat #3025; CST, Danvers, MA, USA), phosphor-insulin receptor substrate 1 (pIRS1; Cat #2381; CST, Danvers, MA, USA), insulin receptor substrate 1 (IRS1; Cat #2382; CST, Danvers, MA, USA), phosphoprotein kinase B (pAkt; Cat #9275; CST, Danvers, MA, USA), protein kinase B (Akt; Cat #9272; CST, Danvers, MA, USA), Akt substrate 160 kDa (AS160; Cat #2670; CST, Danvers, MA, USA), GLUT4 (Cat#2213; CST, Danvers, MA, USA), mTOR (Cat #2972; CST, Danvers, MA, USA), LC3 A/B (Cat #12741; CST, Danvers, MA, USA), uncoupling protein-1 (UCP1; Cat #14670; CST, Danvers, MA, USA), PPARg coactivator 1 alpha (PGC1a; Cat #M00236; Pleasanton, CA, USA), GAPDH (Cat #E11-12057; EnoGene; New York, NY, USA), and vinculin (Cat #4650; CST, Danvers, MA, USA). ..

    Binding Assay:

    Article Title: Gene regulation of RMR-related DNAJC6 on adipogenesis and mitochondria function in 3T3-L1 preadipocytes.
    Article Snippet: .. Reactions with the following primary antibodies (diluted 1:500e1:2000) were performed overnight on the membrane at 4 C: DNAJC6 (Cat #PA5-26981; Invitrogen, Waltham,MA, USA), Pref-1 (Cat #sc-376,755; Santa Cruz, Dallas, TX, USA), peroxisome proliferator-activated receptor gamma (PPARg; Cat #2435; CST, Danvers, MA, USA), CCAAT/enhancer binding protein beta (C/EBPb; Cat #3082; CST, Danvers, MA, USA), CCAAT/ enhancer binding protein alpha (C/EBPa; Cat #8178; CST, Danvers, MA, USA), adipocyte protein 2 (aP2; Cat #sc-271,529; Santa Cruz, Dallas, TX, USA), phosphohormone-sensitive lipase (pHSL; Cat #4139; CST, Danvers, MA, USA), hormone-sensitive lipase (HSL; Cat #4107; CST, Danvers, MA, USA), leptin receptor (LEPR; Cat #ab5593; Abcam), leptin (Cat #675002; Biolegend, San Diego, CA, USA), adiponectin (Cat #2789; CST, Danvers, MA, USA), insulin receptor b (Cat #3025; CST, Danvers, MA, USA), phosphor-insulin receptor substrate 1 (pIRS1; Cat #2381; CST, Danvers, MA, USA), insulin receptor substrate 1 (IRS1; Cat #2382; CST, Danvers, MA, USA), phosphoprotein kinase B (pAkt; Cat #9275; CST, Danvers, MA, USA), protein kinase B (Akt; Cat #9272; CST, Danvers, MA, USA), Akt substrate 160 kDa (AS160; Cat #2670; CST, Danvers, MA, USA), GLUT4 (Cat#2213; CST, Danvers, MA, USA), mTOR (Cat #2972; CST, Danvers, MA, USA), LC3 A/B (Cat #12741; CST, Danvers, MA, USA), uncoupling protein-1 (UCP1; Cat #14670; CST, Danvers, MA, USA), PPARg coactivator 1 alpha (PGC1a; Cat #M00236; Pleasanton, CA, USA), GAPDH (Cat #E11-12057; EnoGene; New York, NY, USA), and vinculin (Cat #4650; CST, Danvers, MA, USA). ..

    Article Title: Nitro-oleic acid regulates growth factor-induced differentiation of bone marrow-derived macrophages
    Article Snippet: Protein samples were subjected to SDS-polyacrylamide gel electrophoresis separation on a 12.5% acrylamide gel and transferred onto polyvinylidene difluoride (PVDF) membrane (Immobilon-P, Millipore, Billerica, MA, USA). .. After incubation in blocking buffer (5% nonfat dried milk in solution of Tris-buffered saline and Tween 20, TBS-T), membranes were probed with primary antibodies overnight at 4°C; primary antibodies against β-actin (cat. no. sc-47778; Santa Cruz Biotechnology, Dallas, TX, USA), iNOS (cat. no. 610431; BD Transduction Transduction Laboratories, Lexington, KY, USA), Arginase I (cat. no. 9819; Cell Signaling Technology, Danvers, MA, USA), M-CSFR (cat. no. 3152; Cell Signaling Technology), phospho-M-CSFR (Tyr723; cat. no. 3155; Cell Signaling Technology), PI3K p85 (cat. no. 4257; Cell Signaling Technology), phospho-PI3K p85/p55 (Tyr458/Tyr199; cat. no. 4228; Cell Signaling Technology), STAT5 (cat. no. 9358; Cell Signaling Technology), phospho-STAT5 (Tyr694; cat. no. 9359; Cell Signaling Technology), p44/42 MAPK (ERK1/2) (cat. no. 4695; Cell Signaling Technology), phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204; cat. no. 4370; Cell Signaling Technology), PU.1 (cat. no. 2266; Cell Signaling Technology), CCAAT/enhancer binding protein beta (C/EBPβ; cat. no. 3087; Cell Signaling Technology), and Avian myelocytomatosis virus oncogene cellular homolog (c-Myc; cat. no. 9402; Cell Signaling Technology) were used. .. Following incubation, membranes were washed three times in TBS-T and then incubated with secondary anti-mouse or anti-rabbit antibodies conjugated with horseradish peroxidase (Cell Signaling Technology) for an hour at room temperature.

    Incubation:

    Article Title: Nitro-oleic acid regulates growth factor-induced differentiation of bone marrow-derived macrophages
    Article Snippet: Protein samples were subjected to SDS-polyacrylamide gel electrophoresis separation on a 12.5% acrylamide gel and transferred onto polyvinylidene difluoride (PVDF) membrane (Immobilon-P, Millipore, Billerica, MA, USA). .. After incubation in blocking buffer (5% nonfat dried milk in solution of Tris-buffered saline and Tween 20, TBS-T), membranes were probed with primary antibodies overnight at 4°C; primary antibodies against β-actin (cat. no. sc-47778; Santa Cruz Biotechnology, Dallas, TX, USA), iNOS (cat. no. 610431; BD Transduction Transduction Laboratories, Lexington, KY, USA), Arginase I (cat. no. 9819; Cell Signaling Technology, Danvers, MA, USA), M-CSFR (cat. no. 3152; Cell Signaling Technology), phospho-M-CSFR (Tyr723; cat. no. 3155; Cell Signaling Technology), PI3K p85 (cat. no. 4257; Cell Signaling Technology), phospho-PI3K p85/p55 (Tyr458/Tyr199; cat. no. 4228; Cell Signaling Technology), STAT5 (cat. no. 9358; Cell Signaling Technology), phospho-STAT5 (Tyr694; cat. no. 9359; Cell Signaling Technology), p44/42 MAPK (ERK1/2) (cat. no. 4695; Cell Signaling Technology), phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204; cat. no. 4370; Cell Signaling Technology), PU.1 (cat. no. 2266; Cell Signaling Technology), CCAAT/enhancer binding protein beta (C/EBPβ; cat. no. 3087; Cell Signaling Technology), and Avian myelocytomatosis virus oncogene cellular homolog (c-Myc; cat. no. 9402; Cell Signaling Technology) were used. .. Following incubation, membranes were washed three times in TBS-T and then incubated with secondary anti-mouse or anti-rabbit antibodies conjugated with horseradish peroxidase (Cell Signaling Technology) for an hour at room temperature.

    Blocking Assay:

    Article Title: Nitro-oleic acid regulates growth factor-induced differentiation of bone marrow-derived macrophages
    Article Snippet: Protein samples were subjected to SDS-polyacrylamide gel electrophoresis separation on a 12.5% acrylamide gel and transferred onto polyvinylidene difluoride (PVDF) membrane (Immobilon-P, Millipore, Billerica, MA, USA). .. After incubation in blocking buffer (5% nonfat dried milk in solution of Tris-buffered saline and Tween 20, TBS-T), membranes were probed with primary antibodies overnight at 4°C; primary antibodies against β-actin (cat. no. sc-47778; Santa Cruz Biotechnology, Dallas, TX, USA), iNOS (cat. no. 610431; BD Transduction Transduction Laboratories, Lexington, KY, USA), Arginase I (cat. no. 9819; Cell Signaling Technology, Danvers, MA, USA), M-CSFR (cat. no. 3152; Cell Signaling Technology), phospho-M-CSFR (Tyr723; cat. no. 3155; Cell Signaling Technology), PI3K p85 (cat. no. 4257; Cell Signaling Technology), phospho-PI3K p85/p55 (Tyr458/Tyr199; cat. no. 4228; Cell Signaling Technology), STAT5 (cat. no. 9358; Cell Signaling Technology), phospho-STAT5 (Tyr694; cat. no. 9359; Cell Signaling Technology), p44/42 MAPK (ERK1/2) (cat. no. 4695; Cell Signaling Technology), phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204; cat. no. 4370; Cell Signaling Technology), PU.1 (cat. no. 2266; Cell Signaling Technology), CCAAT/enhancer binding protein beta (C/EBPβ; cat. no. 3087; Cell Signaling Technology), and Avian myelocytomatosis virus oncogene cellular homolog (c-Myc; cat. no. 9402; Cell Signaling Technology) were used. .. Following incubation, membranes were washed three times in TBS-T and then incubated with secondary anti-mouse or anti-rabbit antibodies conjugated with horseradish peroxidase (Cell Signaling Technology) for an hour at room temperature.

    Saline:

    Article Title: Nitro-oleic acid regulates growth factor-induced differentiation of bone marrow-derived macrophages
    Article Snippet: Protein samples were subjected to SDS-polyacrylamide gel electrophoresis separation on a 12.5% acrylamide gel and transferred onto polyvinylidene difluoride (PVDF) membrane (Immobilon-P, Millipore, Billerica, MA, USA). .. After incubation in blocking buffer (5% nonfat dried milk in solution of Tris-buffered saline and Tween 20, TBS-T), membranes were probed with primary antibodies overnight at 4°C; primary antibodies against β-actin (cat. no. sc-47778; Santa Cruz Biotechnology, Dallas, TX, USA), iNOS (cat. no. 610431; BD Transduction Transduction Laboratories, Lexington, KY, USA), Arginase I (cat. no. 9819; Cell Signaling Technology, Danvers, MA, USA), M-CSFR (cat. no. 3152; Cell Signaling Technology), phospho-M-CSFR (Tyr723; cat. no. 3155; Cell Signaling Technology), PI3K p85 (cat. no. 4257; Cell Signaling Technology), phospho-PI3K p85/p55 (Tyr458/Tyr199; cat. no. 4228; Cell Signaling Technology), STAT5 (cat. no. 9358; Cell Signaling Technology), phospho-STAT5 (Tyr694; cat. no. 9359; Cell Signaling Technology), p44/42 MAPK (ERK1/2) (cat. no. 4695; Cell Signaling Technology), phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204; cat. no. 4370; Cell Signaling Technology), PU.1 (cat. no. 2266; Cell Signaling Technology), CCAAT/enhancer binding protein beta (C/EBPβ; cat. no. 3087; Cell Signaling Technology), and Avian myelocytomatosis virus oncogene cellular homolog (c-Myc; cat. no. 9402; Cell Signaling Technology) were used. .. Following incubation, membranes were washed three times in TBS-T and then incubated with secondary anti-mouse or anti-rabbit antibodies conjugated with horseradish peroxidase (Cell Signaling Technology) for an hour at room temperature.

    Transduction:

    Article Title: Nitro-oleic acid regulates growth factor-induced differentiation of bone marrow-derived macrophages
    Article Snippet: Protein samples were subjected to SDS-polyacrylamide gel electrophoresis separation on a 12.5% acrylamide gel and transferred onto polyvinylidene difluoride (PVDF) membrane (Immobilon-P, Millipore, Billerica, MA, USA). .. After incubation in blocking buffer (5% nonfat dried milk in solution of Tris-buffered saline and Tween 20, TBS-T), membranes were probed with primary antibodies overnight at 4°C; primary antibodies against β-actin (cat. no. sc-47778; Santa Cruz Biotechnology, Dallas, TX, USA), iNOS (cat. no. 610431; BD Transduction Transduction Laboratories, Lexington, KY, USA), Arginase I (cat. no. 9819; Cell Signaling Technology, Danvers, MA, USA), M-CSFR (cat. no. 3152; Cell Signaling Technology), phospho-M-CSFR (Tyr723; cat. no. 3155; Cell Signaling Technology), PI3K p85 (cat. no. 4257; Cell Signaling Technology), phospho-PI3K p85/p55 (Tyr458/Tyr199; cat. no. 4228; Cell Signaling Technology), STAT5 (cat. no. 9358; Cell Signaling Technology), phospho-STAT5 (Tyr694; cat. no. 9359; Cell Signaling Technology), p44/42 MAPK (ERK1/2) (cat. no. 4695; Cell Signaling Technology), phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204; cat. no. 4370; Cell Signaling Technology), PU.1 (cat. no. 2266; Cell Signaling Technology), CCAAT/enhancer binding protein beta (C/EBPβ; cat. no. 3087; Cell Signaling Technology), and Avian myelocytomatosis virus oncogene cellular homolog (c-Myc; cat. no. 9402; Cell Signaling Technology) were used. .. Following incubation, membranes were washed three times in TBS-T and then incubated with secondary anti-mouse or anti-rabbit antibodies conjugated with horseradish peroxidase (Cell Signaling Technology) for an hour at room temperature.

    Virus:

    Article Title: Nitro-oleic acid regulates growth factor-induced differentiation of bone marrow-derived macrophages
    Article Snippet: Protein samples were subjected to SDS-polyacrylamide gel electrophoresis separation on a 12.5% acrylamide gel and transferred onto polyvinylidene difluoride (PVDF) membrane (Immobilon-P, Millipore, Billerica, MA, USA). .. After incubation in blocking buffer (5% nonfat dried milk in solution of Tris-buffered saline and Tween 20, TBS-T), membranes were probed with primary antibodies overnight at 4°C; primary antibodies against β-actin (cat. no. sc-47778; Santa Cruz Biotechnology, Dallas, TX, USA), iNOS (cat. no. 610431; BD Transduction Transduction Laboratories, Lexington, KY, USA), Arginase I (cat. no. 9819; Cell Signaling Technology, Danvers, MA, USA), M-CSFR (cat. no. 3152; Cell Signaling Technology), phospho-M-CSFR (Tyr723; cat. no. 3155; Cell Signaling Technology), PI3K p85 (cat. no. 4257; Cell Signaling Technology), phospho-PI3K p85/p55 (Tyr458/Tyr199; cat. no. 4228; Cell Signaling Technology), STAT5 (cat. no. 9358; Cell Signaling Technology), phospho-STAT5 (Tyr694; cat. no. 9359; Cell Signaling Technology), p44/42 MAPK (ERK1/2) (cat. no. 4695; Cell Signaling Technology), phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204; cat. no. 4370; Cell Signaling Technology), PU.1 (cat. no. 2266; Cell Signaling Technology), CCAAT/enhancer binding protein beta (C/EBPβ; cat. no. 3087; Cell Signaling Technology), and Avian myelocytomatosis virus oncogene cellular homolog (c-Myc; cat. no. 9402; Cell Signaling Technology) were used. .. Following incubation, membranes were washed three times in TBS-T and then incubated with secondary anti-mouse or anti-rabbit antibodies conjugated with horseradish peroxidase (Cell Signaling Technology) for an hour at room temperature.



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    αPD-L1 inhibits TAM/M2 polarization via the prevention of STAT3 phosphorylation and nuclear translocation. (A) The protein expression levels of CEBPβ, p-ERK, ERK, p-STAT6, STAT6, p-STAT3 and STAT3 in RAW264.7 cells treated with IL-13 and/or αPD-L1 were determined using western blot analysis. (B and C) The protein expression levels and nuclear translocation of STAT3 and p-STAT3 in RAW264.7 cells treated with IL-13 and/or αPD-L1 were assessed using immunofluorescence staining (magnification, ×400). (D) Cytosol and nuclear protein expression levels of STAT3 and p-STAT3 in RAW264.7 cells treated with IL-13 and/or αPD-L1 were assessed using western blot analysis. * P<0.05, ** P<0.01 and *** P<0.001. ns, not significant; αPD-L1, programmed death-ligand 1 inhibitor; TAM/M2, tumor-associated macrophages/M2-type; CEBPβ, cAMP response <t>element-binding</t> <t>protein/CCAAT-enhancer-binding</t> protein β; p-, phosphorylated.
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    Figure 6. αPD‑L1 inhibits TAM/M2 polarization via the prevention of STAT3 phosphorylation and nuclear translocation. (A) The protein expression levels of <t>CEBPβ,</t> p‑ERK, ERK, p‑STAT6, STAT6, p‑STAT3 and STAT3 in RAW264.7 cells treated with IL‑13 and/or αPD‑L1 were determined using western blot analysis. (B and C) The protein expression levels and nuclear translocation of STAT3 and p‑STAT3 in RAW264.7 cells treated with IL‑13 and/or αPD‑L1 were assessed using immunofluorescence staining (magnification, x400). (D) Cytosol and nuclear protein expression levels of STAT3 and p‑STAT3 in RAW264.7 cells treated with IL‑13 and/or αPD‑L1 were assessed using western blot analysis. *P<0.05, **P<0.01 and ***P<0.001. ns, not significant; αPD‑L1, programmed death‑ligand 1 inhibitor; TAM/M2, tumor‑associated macrophages/M2‑type; CEBPβ, cAMP response element‑binding protein/CCAAT‑enhancer‑binding protein β; p‑, phosphorylated.
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    Image Search Results


    Genes expression in mammary glands at 130 days of age.

    Journal: BMC Cancer

    Article Title: Maternal consumption of canola oil suppressed mammary gland tumorigenesis in C3(1) TAg mice offspring

    doi: 10.1186/1471-2407-10-81

    Figure Lengend Snippet: Genes expression in mammary glands at 130 days of age.

    Article Snippet: The blots were blocked with 5% BSA in TBST overnight at 4°C and probed with primary antibodies against [CCAAT-enhancer binding proteins β (C/EBPβ) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA), fatty acid synthase (Santa Cruz Biotechnology, Inc., Santa Cruz, CA), glyceraldehyde 3 phosphate dehydrogenase (GAPDH) (CHEMICON International, Billerica, MA) or cytokeratin 8 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA,)] in blocking buffer for 1 hour at room temperature.

    Techniques: Expressing, Binding Assay, Transformation Assay

    Results of protein analyses for fatty acid synthase (Fas) and CCAAT/enhancer binding protein (C/EBP), beta at 130 days of age . Western blot for fatty acid synthase showed that Fas protein was significantly less (p = 0.02 by T-test, n = 3/group) and that C/EBP, β was not quite significantly higher (p = 0.06 by T-Test, n = 3/group) in mammary glands of CA/CO mice than in CO/CO mice at 130 days. Values shown are relative density and have been corrected for protein loading (GAPDH) and for cytokeratin (to correct for the epithelial compartment of the assayed sample).

    Journal: BMC Cancer

    Article Title: Maternal consumption of canola oil suppressed mammary gland tumorigenesis in C3(1) TAg mice offspring

    doi: 10.1186/1471-2407-10-81

    Figure Lengend Snippet: Results of protein analyses for fatty acid synthase (Fas) and CCAAT/enhancer binding protein (C/EBP), beta at 130 days of age . Western blot for fatty acid synthase showed that Fas protein was significantly less (p = 0.02 by T-test, n = 3/group) and that C/EBP, β was not quite significantly higher (p = 0.06 by T-Test, n = 3/group) in mammary glands of CA/CO mice than in CO/CO mice at 130 days. Values shown are relative density and have been corrected for protein loading (GAPDH) and for cytokeratin (to correct for the epithelial compartment of the assayed sample).

    Article Snippet: The blots were blocked with 5% BSA in TBST overnight at 4°C and probed with primary antibodies against [CCAAT-enhancer binding proteins β (C/EBPβ) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA), fatty acid synthase (Santa Cruz Biotechnology, Inc., Santa Cruz, CA), glyceraldehyde 3 phosphate dehydrogenase (GAPDH) (CHEMICON International, Billerica, MA) or cytokeratin 8 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA,)] in blocking buffer for 1 hour at room temperature.

    Techniques: Binding Assay, Western Blot

    αPD-L1 inhibits TAM/M2 polarization via the prevention of STAT3 phosphorylation and nuclear translocation. (A) The protein expression levels of CEBPβ, p-ERK, ERK, p-STAT6, STAT6, p-STAT3 and STAT3 in RAW264.7 cells treated with IL-13 and/or αPD-L1 were determined using western blot analysis. (B and C) The protein expression levels and nuclear translocation of STAT3 and p-STAT3 in RAW264.7 cells treated with IL-13 and/or αPD-L1 were assessed using immunofluorescence staining (magnification, ×400). (D) Cytosol and nuclear protein expression levels of STAT3 and p-STAT3 in RAW264.7 cells treated with IL-13 and/or αPD-L1 were assessed using western blot analysis. * P<0.05, ** P<0.01 and *** P<0.001. ns, not significant; αPD-L1, programmed death-ligand 1 inhibitor; TAM/M2, tumor-associated macrophages/M2-type; CEBPβ, cAMP response element-binding protein/CCAAT-enhancer-binding protein β; p-, phosphorylated.

    Journal: International Journal of Oncology

    Article Title: PD-L1 mediates triple-negative breast cancer evolution via the regulation of TAM/M2 polarization

    doi: 10.3892/ijo.2022.5440

    Figure Lengend Snippet: αPD-L1 inhibits TAM/M2 polarization via the prevention of STAT3 phosphorylation and nuclear translocation. (A) The protein expression levels of CEBPβ, p-ERK, ERK, p-STAT6, STAT6, p-STAT3 and STAT3 in RAW264.7 cells treated with IL-13 and/or αPD-L1 were determined using western blot analysis. (B and C) The protein expression levels and nuclear translocation of STAT3 and p-STAT3 in RAW264.7 cells treated with IL-13 and/or αPD-L1 were assessed using immunofluorescence staining (magnification, ×400). (D) Cytosol and nuclear protein expression levels of STAT3 and p-STAT3 in RAW264.7 cells treated with IL-13 and/or αPD-L1 were assessed using western blot analysis. * P<0.05, ** P<0.01 and *** P<0.001. ns, not significant; αPD-L1, programmed death-ligand 1 inhibitor; TAM/M2, tumor-associated macrophages/M2-type; CEBPβ, cAMP response element-binding protein/CCAAT-enhancer-binding protein β; p-, phosphorylated.

    Article Snippet: The membranes were incubated with the primary antibodies at 4°C overnight and subsequently with the secondary antibodies at room temperature for 1 h. Primary antibodies specific for the following proteins were used: PD-L1 (1:1,000; cat. no. 60475 and 85164; CST Biological Reagents Co., Ltd.), CD86 (1:1,000; cat. no. sc-28347; Santa Cruz Biotechnology, Inc.), CD206 (1:1,000; cat. no. 24595; CST Biological Reagents Co., Ltd.), VEGF (1:1,000; cat. no. sc-7269; Santa Cruz Biotechnology, Inc.), MMP2 (1:1,000; cat. no. sc-13594; Santa Cruz Biotechnology, Inc.), MMP9 (1:1,000; cat. no. sc-393859; Santa Cruz Biotechnology, Inc.), E-cadherin (1:1,000; cat. no. 14472; CST Biological Reagents Co., Ltd.), zonula occludens-1 (ZO-1; 1:1,000; cat. no. 8193; CST Biological Reagents Co., Ltd.), N-cadherin (1:1,000; cat. no. sc-8424; Santa Cruz Biotechnology, Inc.), vimentin (1:1,000; cat. no. sc-6260; Santa Cruz Biotechnology, Inc.), Slug (1:1,000; cat. no. sc-166476; Santa Cruz Biotechnology, Inc.), Twist (1:1,000; cat. no. sc-81417; Santa Cruz Biotechnology, Inc.), CD44 (1:1,000; cat. no. 3570; CST Biological Reagents Co., Ltd.), octamer-binding transcription factor 4 (Oct4; 1:1,000; cat. no. 75643; CST Biological Reagents Co., Ltd.), Nanog (1:1,000; cat. no. sc-374103; Santa Cruz Biotechnology, Inc.), Bmi1 (1:1,000; cat. no. sc-390443; Santa Cruz Biotechnology, Inc.), Sox2 (1:1,000; cat. no. sc-365823; Santa Cruz Biotechnology, Inc.), CCAAT/enhancer-binding protein β (CEBPβ; 1:1,000; cat. no. sc-7962; Santa Cruz Biotechnology, Inc.), phosphorylated (p-)ERK (1:1,000; cat. no. 4695; CST Biological Reagents Co., Ltd.), ERK (1:1,000; cat. no. 48303; CST Biological Reagents Co., Ltd.), p-STAT6 (1:1,000; cat. no. 56554; CST Biological Reagents Co., Ltd.), STAT6 (1:1,000; cat. no. sc-374021; Santa Cruz Biotechnology, Inc.), p-STAT3 (1:1,000; cat. no. 9145; CST Biological Reagents Co., Ltd.), STAT3 (1:1,000; cat. no. 9139; CST Biological Reagents Co., Ltd.), β-actin (1:1,000; cat. no. CW0096; CWBio Technology Co., Ltd.), GAPDH (1:1,000; cat. no. CW0100M; CWBio Technology Co., Ltd.) and H3 (1:1,000; cat. no. 60932; CST Biological Reagents Co., Ltd.).

    Techniques: Translocation Assay, Expressing, Western Blot, Immunofluorescence, Staining, Binding Assay

    Figure 6. αPD‑L1 inhibits TAM/M2 polarization via the prevention of STAT3 phosphorylation and nuclear translocation. (A) The protein expression levels of CEBPβ, p‑ERK, ERK, p‑STAT6, STAT6, p‑STAT3 and STAT3 in RAW264.7 cells treated with IL‑13 and/or αPD‑L1 were determined using western blot analysis. (B and C) The protein expression levels and nuclear translocation of STAT3 and p‑STAT3 in RAW264.7 cells treated with IL‑13 and/or αPD‑L1 were assessed using immunofluorescence staining (magnification, x400). (D) Cytosol and nuclear protein expression levels of STAT3 and p‑STAT3 in RAW264.7 cells treated with IL‑13 and/or αPD‑L1 were assessed using western blot analysis. *P<0.05, **P<0.01 and ***P<0.001. ns, not significant; αPD‑L1, programmed death‑ligand 1 inhibitor; TAM/M2, tumor‑associated macrophages/M2‑type; CEBPβ, cAMP response element‑binding protein/CCAAT‑enhancer‑binding protein β; p‑, phosphorylated.

    Journal: International journal of oncology

    Article Title: PD‑L1 mediates triple‑negative breast cancer evolution via the regulation of TAM/M2 polarization.

    doi: 10.3892/ijo.2022.5440

    Figure Lengend Snippet: Figure 6. αPD‑L1 inhibits TAM/M2 polarization via the prevention of STAT3 phosphorylation and nuclear translocation. (A) The protein expression levels of CEBPβ, p‑ERK, ERK, p‑STAT6, STAT6, p‑STAT3 and STAT3 in RAW264.7 cells treated with IL‑13 and/or αPD‑L1 were determined using western blot analysis. (B and C) The protein expression levels and nuclear translocation of STAT3 and p‑STAT3 in RAW264.7 cells treated with IL‑13 and/or αPD‑L1 were assessed using immunofluorescence staining (magnification, x400). (D) Cytosol and nuclear protein expression levels of STAT3 and p‑STAT3 in RAW264.7 cells treated with IL‑13 and/or αPD‑L1 were assessed using western blot analysis. *P<0.05, **P<0.01 and ***P<0.001. ns, not significant; αPD‑L1, programmed death‑ligand 1 inhibitor; TAM/M2, tumor‑associated macrophages/M2‑type; CEBPβ, cAMP response element‑binding protein/CCAAT‑enhancer‑binding protein β; p‑, phosphorylated.

    Article Snippet: The membranes were incubated with the primary antibodies at 4 ̊C overnight and subsequently with the secondary antibodies at room temperature for 1 h. Primary antibodies specific for the following proteins were used: PD‐L1 (1:1,000; cat. no. 60475 and 85164; CST Biological Reagents Co., Ltd.), CD86 (1:1,000; cat. no. sc‐28347; Santa Cruz Biotechnology, Inc.), CD206 (1:1,000; cat. no. 24595; CST Biological Reagents Co., Ltd.), VEGF (1:1,000; cat. no. sc‐7269; Santa Cruz Biotechnology, Inc.), MMP2 (1:1,000; cat. no. sc‐13594; Santa Cruz Biotechnology, Inc.), MMP9 (1:1,000; cat. no. sc‐393859; Santa Cruz Biotechnology, Inc.), E‐cadherin (1:1,000; cat. no. 14472; CST Biological Reagents Co., Ltd.), zonula occludens‐1 (ZO‐1; 1:1,000; cat. no. 8193; CST Biological Reagents Co., Ltd.), N‐cadherin (1:1,000; cat. no. sc‐8424; Santa Cruz Biotechnology, Inc.), vimentin (1:1,000; cat. no. sc‐6260; Santa Cruz Biotechnology, Inc.), Slug (1:1,000; cat. no. sc‐166476; Santa Cruz Biotechnology, Inc.), Twist (1:1,000; cat. no. sc‐81417; Santa Cruz Biotechnology, Inc.), CD44 (1:1,000; cat. no. 3570; CST Biological Reagents Co., Ltd.), octamer‐binding transcription factor 4 (Oct4; 1:1,000; cat. no. 75643; CST Biological Reagents Co., Ltd.), Nanog (1:1,000; cat. no. sc‐374103; Santa Cruz Biotechnology, Inc.), Bmi1 (1:1,000; cat. no. sc‐390443; Santa Cruz Biotechnology, Inc.), Sox2 (1:1,000; cat. no. sc‐365823; Santa Cruz Biotechnology, Inc.), CCAAT/enhancer‐binding protein β (CEBPβ; 1:1,000; cat. no. sc‐7962; Santa Cruz Biotechnology, Inc.), phosphorylated (p‐)ERK (1:1,000; cat. no. 4695; CST Biological Reagents Co., Ltd.), ERK (1:1,000; cat. no. 48303; CST Biological Reagents Co., Ltd.), p‐STAT6 (1:1,000; cat. no. 56554; CST Biological Reagents Co., Ltd.), STAT6 (1:1,000; cat. no. sc‐374021; Santa Cruz Biotechnology, Inc.), p‐STAT3 (1:1,000; cat. no. 9145; CST Biological Reagents Co., Ltd.), STAT3 (1:1,000; cat. no. 9139; CST Biological Reagents Co., Ltd.), β‐actin (1:1,000; cat. no. CW0096; CWBio Technology Co., Ltd.), GAPDH (1:1,000; cat. no. CW0100M; CWBio Technology Co., Ltd.) and H3 (1:1,000; cat. no. 60932; CST Biological Reagents Co., Ltd.).

    Techniques: Phospho-proteomics, Translocation Assay, Expressing, Western Blot, Immunofluorescence, Staining