- 12-7209-41 - Invitrogen Antibodies CSF2 antibody

Lin J, Tang J, Lin J, He Y, Yu Z, Jiang R, Yang B, Ou Q
JCI insight 2021 Nov 22;6(22)

Shi Y, Liu CH, Roberts AI, Das J, Xu G, Ren G, Zhang Y, Zhang L, Yuan ZR, Tan HS, Das G, Devadas S
Cell research 2006 Feb;16(2):126-33

Metcalf D
Blood 1986 Feb;67(2):257-67

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Normal human peripheral blood mononuclear cells were cultured 5 hours with Protein Transport Inhibitor Cocktail (Product # 00-4980-03) (unstimulated) (left) or with Cell Stimulation Cocktail (Product # 00-4975-03) (right). The cells were stained intracellularly with Anti-Human CD4 APC (Product # 17-0047-42) and Anti-Human GM-CSF PE using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Cells in the lymphocyte gate were used for analysis.
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Increased pTh17 cells have potential relationship with YY1 expression in RA. ( A and B ) Representative images of flow cytometry results for the proportion of CD4 + IL-17A + IFN-gamma + cells and CD4 + IL-17A + GM-CSF + cells in PBMCs of patients with RA and OA as well as HDs. Data presented as mean +- SD ( n = 11). * P < 0.05 (ANOVA). ( C ) The relative gene expression levels of YY1 in PBMCs of patients with RA ( n = 33) and OA ( n = 24) as well as HDs ( n = 32). Data presented as box-and-whisker plot (The line within the box and the bounds of the box represent median and interquartile range, respectively. The whiskers denote the 25th percentile minus 1.5 interquartile range and 75th percentile plus 1.5 interquartile range. The points, which are outside the whisker, represent outliers). *** P < 0.001, **** P < 0.0001 (ANOVA). ( D and E ) Analysis of the correlation of IL-17A or IL-22 mRNA and YY1 mRNA in PBMCs of RA patients ( n = 41, Pearson correlation). ( F ) The relative gene expression levels of YY1 in purified CD4 + T cells of patients with RA ( n = 36) and HD ( n = 18). Data presented as box-and-whisker plot (The meaning of the symbols is the same as C ). *** P < 0.001 (Student's t test). ( G and H ) Analysis of the correlation of IL-17A or IL-22 mRNA and YY1 mRNA in purified CD4 + T cells of RA patients ( n = 36, Pearson correlation).
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 YY1 is involved in pTh17 cell differentiation. ( A ) Representative Western blots of YY1 in an ex vivo Th17 subset differentiation system including unpolarized, non-pTh17-polarized, and pTh17-polarized groups (at least 3 independent experiments). ( B ) Representative fluorescence images for transfection efficiency of the sh-YY1 lentivirus (scale bars = 100 mum, at least 3 independent experiments). ( C ) The relative expression levels of YY1 for knockdown effect of the sh-YY1 lentivirus ( n = 3 independent experiments). ( D ) Representative Western blots of YY1 for knockdown effect of the sh-YY1 lentivirus (at least 3 independent experiments). ( E and F ) Representative images of flow cytometry results for the proportion of CD4 + IL-17A + IFN-gamma + cells and CD4 + IL-17A + GM-CSF + cells in the pTh17-polarized group treated with the sh-YY1 lentivirus. All data presented as mean +- SEM ( n = 3 of independent experiments). ** P < 0.01, * P < 0.05 (ANOVA).
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 YY1 specifically regulates pTh17 cell differentiation through affecting T-bet. ( A ) Heatmap of clustering analysis of differentially expressed genes of cells treated with the sh-YY1 lentiviruses in pTh17-polarized conditions. ( B ) The relative gene expressions of pTh17-related transcription factors and cytokines in cells from YY1-knockdown pTh17-polarized group. Data presented as mean +- SEM ( n = 6). *** P < 0.001, **** P < 0.0001 (Student's t test). ( C ) Representative Western blots of T-bet in an ex vivo Th17 subset differentiation system including unpolarized, non-pTh17-polarized, and pTh17-polarized (at least 3 independent experiments). ( D and E ) Representative images of flow cytometry results for the proportion of CD4 + IL-17A + IFN-gamma + cells and CD4 + IL-17A + GM-CSF + cells in pTh17-polarized group treated with the sh-YY1 lentivirus or cotransfected with sh-YY1 lentivirus and T-bet-overexpressing lentivirus. Data presented as mean +- SEM ( n = 3 independent experiments). ** P < 0.01, * P < 0.05 (ANOVA). ( F and G ) Representative images of flow cytometry results for the proportion of CD4 + IL-17A + IFN-gamma + cells in Th1-polarized group treated with the sh-YY1 lentivirus. Data presented as mean +- SEM ( n = 3 independent experiments). * P < 0.05 (ANOVA).
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 7 miR-124-3p is involved in the pathogenicity of Th17 cells. ( A and B ) Representative images of flow cytometry results for the proportion of CD4 + IL-17A + IFN-gamma + cells and CD4 + IL-17A + GM-CSF + cells in pTh17-polarized group transfected with miR-124-3p mimics, inhibitors, or control plasmid. Data presented as mean +- SEM ( n = 3 independent experiments). * P < 0.05 (ANOVA). ( C - F ) Analysis of the correlation of miR-124-3p and anti-CCP, RF, CRP, and ESR in purified CD4 + T cells of patients with RA ( n = 36. C , E , and F using Pearson correlation, D using Spearman correlation). ( G ) Sketch of miR-124-3p and YY1 in differentiation of pTh17 cells in RA.
Conjugate: Yellow dye

12-7209-41