anti epor rabbit polyclonal  (Bioss)

Journal: eLife

Article Title: Differential regulation by CD47 and thrombospondin-1 of extramedullary erythropoiesis in mouse spleen

doi: 10.7554/eLife.92679

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-EPOR (Rabbit polyclonal) , Bioss; Thermo Fisher , Cat#: BS-1424R , FACS (1 μg/100 μl/1 million cells).

Techniques: Cell Isolation, Recombinant, Control

Journal: Frontiers in Immunology

Article Title: Extracellular CIRP-Impaired Rab26 Restrains EPOR-Mediated Macrophage Polarization in Acute Lung Injury

doi: 10.3389/fimmu.2021.768435

Figure Lengend Snippet: Myeloid EPOR signaling is essential for macrophage polarization. (A) A qPCR assay was conducted to evaluate the mRNA expression of EPOR and PPARγ after WT BMDMs were treated with eCIRP for the indicated times (n = 3). (B) The MFIs of cell surface EPOR in BMDMs treated with eCIRP for the indicated times were tested by FACS (n = 3). ** P < 0.01 vs. 0 h group. (C) The percentage of EPOR + cells in WT and EPOR cKO BMDMs treated with or without eCIRP for 24 h (n = 3). (D) The MFIs of CD80 and CD86 in WT and EPOR cKO BMDMs were measured by FACS with or without eCIRP (1 µg/ml) and rhEPO (20 IU/ml) administration (n = 3). (E) A qPCR assay was conducted to evaluate the mRNA expression of TNF-α, IL-6, and IL-1β in WT and EPOR cKO BMDMs with or without eCIRP and rhEPO (20 IU/ml) administration (n = 3). (F) The supernatant concentrations of TNF-α, IL-6, and IL-10 in BMDMs were measured with or without eCIRP (1 µg/ml) and rhEPO (20 IU/ml) administration (n = 3). (G) Western blot analysis of PPARγ and β-actin protein expression was conducted after WT and EPOR cKO BMDMs were treated with eCIRP for 24 h (n = 3). Data are representative of at least two independent experiments. Results were expressed as mean ± SD. n.s., not statistically significant. * P < 0.05, ** P < 0.01. Statistics: One-way ANOVA with Tukey’s post-hoc test for multiple comparisons (A–G) . EPOR, erythropoietin receptor; PCR, Polymerase Chain Reaction; PPAR, peroxisome proliferator-activated receptor; BMDM, bone marrow derived macrophage; eCIRP, extracellular cold-inducible RNA-binding protein; WT, wild type; EPO, erythropoietin; MFI, mean fluorescence intensity; FACS, fluorescence activated cell sorter.

Article Snippet: Antibodies were as follows: APC-labeled anti-mouse F4/80 antibody (BioLegend, #123116, clone #BM8), APC/Cyanine7-labeled anti-mouse F4/80 antibody (BioLegend, #123118, clone #BM8), PE-labeled anti-mouse F4/80 antibody (BioLegend, #123110, clone #BM8), PE-labeled anti-mouse Ly-6G antibody (BioLegend, #127608, clone #1A8), APC/Cyanine7-labeled anti-mouse Ly-6G antibody (BioLegend, #127624, clone #1A8), APC/Cyanine7-labeled anti-mouse/human CD11b antibody (BioLegend, #101226, clone #M1/70), APC-labeled anti-mouse TNF-α antibody (BioLegend, #506308, clone #MP6-XT22), APC-labeled anti-mouse IL-6 antibody (BioLegend, #504508, clone #MP5-20F3), APC-labeled anti-mouse IL-10 antibody (BioLegend, #505010, clone #JES5-16E3), PE-labeled anti-mouse EPOR antibody (Bioss, #bs-1424R), FITC-labeled anti-mouse CD86 (BD, #553691, clone #GL-1), APC-labeled anti-mouse CD86 (BioLegend, #105012, clone #GL-1), PE-labeled anti-mouse CD80 (eBioscience, #12-0801-81, clone #16-10A1), PE/Cyanine7-labeled anti-mouse CD80 (BioLegend, #104734, clone #16-10A1), APC-labeled anti-mouse CD206 (BioLegend, #141707, clone #C068C2), PE-labeled anti-mouse CD206 (BioLegend, #141706, clone #C068C2), eBioscience™ Fixable Viability Dye eFluor™ 450 (Invitrogen, #65-0863-14), anti-CD16/32 antibody (Sungene Biotech, #M10161-14F); Mouse Erythropoietin R Antibody (RD, #AF1390), anti-PPARγ antibody (CTS, #2430), anti-Rab26 antibody (Abcam, #ab198202), anti-β-Actin antibody (CST, #4970S), Peroxidase AffiniPure Goat Anti-Rabbit IgG (Jackson Immunoresearch, #111-035-003), mouse anti-goat IgG-FITC (Santa Cruz, #sc-2356), and CIRP polyclonal antibody (proteintech, #10209-2-AP).

Techniques: Expressing, Western Blot, Polymerase Chain Reaction, Derivative Assay, RNA Binding Assay, Fluorescence

Journal: Frontiers in Immunology

Article Title: Extracellular CIRP-Impaired Rab26 Restrains EPOR-Mediated Macrophage Polarization in Acute Lung Injury

doi: 10.3389/fimmu.2021.768435

Figure Lengend Snippet: Rab26 deficiency reduces EPOR signaling and restrains macrophage polarization. A qPCR assay was conducted to evaluate the mRNA expression of EPOR (A) and PPARγ (B) in WT BMDMs treated with rhEPO (20 IU/ml) for the indicated time (n = 3). (C) The MFIs of cell surface EPOR in BMDMs treated with rhEPO (20 IU/ml) for the indicated times were tested by FACS (n = 3). * P < 0.05, ** P < 0.01 vs. 0 h. (D) The MFIs of CD80 and CD86 were evaluated by FACS in WT and Rab26 -/- BMDMs treated with or without eCIRP (1 µg/ml) or rhEPO (20 IU/ml) for 24 h (n = 3). (E) A qPCR assay was conducted to evaluate the mRNA expression of TNF-α, IL-6, and IL-1β in WT and Rab26 -/- BMDMs treated with eCIRP (1 µg/ml) and rhEPO (20 IU/ml) for 24 h (n = 3). (F) Localization of EPOR in WT and Rab26 -/- BMDMs. WT and Rab26 -/- BMDMs were stained with an anti-EPOR antibody (1:100 dilution) and Alexa Fluor 488-labeled secondary antibody (1:200 dilution) (green), and nuclei were stained with DAPI (blue). Representative confocal images of the surface and intracellular expression of EPOR are shown. Scale bar: 10 µm. Data are representative of at least two independent experiments. Results were expressed as mean ± SD. n. s., not statistically significant. * P < 0.05, ** P < 0.01. Statistics: One-way ANOVA with Tukey’s post-hoc test for multiple comparisons (A–E) . EPOR, erythropoietin receptor; PCR, polymerase chain reaction; PPAR, peroxisome proliferator-activated receptor; BMDM, bone marrow derived macrophage; EPO, erythropoietin; MFI, mean fluorescence intensity; FACS, fluorescence activated cell sorter; eCIRP, extracellular cold-inducible RNA-binding protein.

Article Snippet: Antibodies were as follows: APC-labeled anti-mouse F4/80 antibody (BioLegend, #123116, clone #BM8), APC/Cyanine7-labeled anti-mouse F4/80 antibody (BioLegend, #123118, clone #BM8), PE-labeled anti-mouse F4/80 antibody (BioLegend, #123110, clone #BM8), PE-labeled anti-mouse Ly-6G antibody (BioLegend, #127608, clone #1A8), APC/Cyanine7-labeled anti-mouse Ly-6G antibody (BioLegend, #127624, clone #1A8), APC/Cyanine7-labeled anti-mouse/human CD11b antibody (BioLegend, #101226, clone #M1/70), APC-labeled anti-mouse TNF-α antibody (BioLegend, #506308, clone #MP6-XT22), APC-labeled anti-mouse IL-6 antibody (BioLegend, #504508, clone #MP5-20F3), APC-labeled anti-mouse IL-10 antibody (BioLegend, #505010, clone #JES5-16E3), PE-labeled anti-mouse EPOR antibody (Bioss, #bs-1424R), FITC-labeled anti-mouse CD86 (BD, #553691, clone #GL-1), APC-labeled anti-mouse CD86 (BioLegend, #105012, clone #GL-1), PE-labeled anti-mouse CD80 (eBioscience, #12-0801-81, clone #16-10A1), PE/Cyanine7-labeled anti-mouse CD80 (BioLegend, #104734, clone #16-10A1), APC-labeled anti-mouse CD206 (BioLegend, #141707, clone #C068C2), PE-labeled anti-mouse CD206 (BioLegend, #141706, clone #C068C2), eBioscience™ Fixable Viability Dye eFluor™ 450 (Invitrogen, #65-0863-14), anti-CD16/32 antibody (Sungene Biotech, #M10161-14F); Mouse Erythropoietin R Antibody (RD, #AF1390), anti-PPARγ antibody (CTS, #2430), anti-Rab26 antibody (Abcam, #ab198202), anti-β-Actin antibody (CST, #4970S), Peroxidase AffiniPure Goat Anti-Rabbit IgG (Jackson Immunoresearch, #111-035-003), mouse anti-goat IgG-FITC (Santa Cruz, #sc-2356), and CIRP polyclonal antibody (proteintech, #10209-2-AP).

Techniques: Expressing, Staining, Labeling, Polymerase Chain Reaction, Derivative Assay, Fluorescence, RNA Binding Assay