Journal: eLife

Article Title: JAK inhibition decreases the autoimmune burden in Down syndrome

doi: 10.7554/eLife.99323

Figure Lengend Snippet:

Article Snippet: Antibody , Mouse monoclonal anti-human CD34 (clone 581) , Fluidigm , Cat # 3163014B; RRID: AB_2811091 , Lot 2651705, 1:33.

Techniques: Isolation, Biomarker Discovery, Staining, Antibody Labeling, Software, Sequencing

Journal: eLife

Article Title: A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro

doi: 10.7554/eLife.94884

Figure Lengend Snippet: ( A ) Single-cell transcriptomic analysis of developing aorta-gonad-mesonephros (AGM) collected from human embryos at Carnegie stages 14 and 15 enriched for CD31 + and CD34 + cells from , Nature. Arterial haemogenic endothelium, dashed line, is identified by the colocalisation of DLL4 and RUNX1 . ( B ) Gene expression analysis of the nine selected target genes in the developing human aorta, showing their expression in the haemogenic endothelium.

Article Snippet: Antibody , Anti-human CD34 PE (mouse monoclonal) , eBioscience , Clone 4H11 , Flow cytometry 1:200.

Techniques: Gene Expression, Expressing

Journal: eLife

Article Title: A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro

doi: 10.7554/eLife.94884

Figure Lengend Snippet: ( A ) Schematic of the derivation of the inducible SAM (iSAM) cell line by ZNFs mediated targeting of the AAVS1 locus and the subsequent derivation of the iSAM_NT (containing the non-targeting control gRNA) and iSAM_AGM (containing the gRNAs for the target genes) by viral transduction of the gRNAs. ( B ) Schematic of the differentiation protocol with activation of the target genes, used for both the control line iSAM_NT and iSAM_AGM. ( C ) Schematic of the IGFBP2 functional validation experiment. ( D ) Expansion of the arterial population marker by membrane expression of DLL4+ following targets’ activation, quantified by flow cytometry at day 8 of differentiation (*p=0.0190, **p=0.0011, Sidak’s two-way ANOVA). ( E ) Flow cytometry analysis of the cell cycle analysis of suspension progenitor cells was obtained following the OP9 coculture of cells treated with IGFBP2 and control at day 13. ( F ) iSAM expressing cells at day 8 of differentiation upon doxycycline (DOX) treatment. ( G ) Target genes’ expression profile across the different libraries, the colour legend for the libraries is on the top right. ( H ) IGFBP2 expression profile across the cell clusters and in the cell lines and conditions indicated in the title (cluster legend same as in I). ( I ) Contribution of each of the libraries to the cell clusters, grouped by libraries (cluster legend at the bottom). Figure 3—figure supplement 1—source data 1. Spreadsheet source file containing the source data used for the plots in . Each tab is labelled to uniquely refer to a specific panel.

Article Snippet: Antibody , Anti-human CD34 PE (mouse monoclonal) , eBioscience , Clone 4H11 , Flow cytometry 1:200.

Techniques: Control, Transduction, Activation Assay, Functional Assay, Biomarker Discovery, Marker, Membrane, Expressing, Flow Cytometry, Cell Cycle Assay, Suspension

Journal: eLife

Article Title: A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro

doi: 10.7554/eLife.94884

Figure Lengend Snippet: ( A ) Gene expression profile of target genes following target genes’ activation, heatmap shows the expression level of the target genes in the iSAM_NT and iSAM_AGM treated with doxycycline (DOX) following normalisation on the -DOX control. ( B ) Dimension reduction and clustering analysis of the scRNAseq data following activation, filtered on cells where the gRNA expression was detected. ( C ) Arterial ( GJA4 , DLL4 ), venous ( NRP2 , APLNR ), and haemogenic marker ( CD44 , RUNX1 ) expression distribution in the clusters indicated by the colour. ( D ) Expression distribution visualised on the UMAP plot showing the location of arterial cells marked by DLL4 , and haemogenic endothelium marked by CD44 and RUNX1 . ( E ) Heatmap of the top 15 marker genes for each of the clusters. ( F ) Contribution of the different libraries to the clusters showing that arterial cell cluster is overrepresented in the iSAM_AGM treated with DOX, compared to the other libraries. ( G ) Expansion of the arterial population assessed by the membrane marker expression of DLL4+ following targets’ activation, quantified by flow cytometry at day 8 of differentiation. (Data are normalised on the iSAM_NT+DOX sample, n=5 independent differentiations, *p=0.0417 paired t-test.) ( H ) Colony-forming potential of the suspension progenitor cells derived from the two lines treated with or without DOX following OP9 coculture activation, data show the colony obtained for 104 CD34+ input equivalent (n=3 from independent differentiations *p<0.05, Tukey’s two-way ANOVA). Figure 3—source data 1. Spreadsheet source file containing the source data used for the plots in . Each tab is labelled to uniquely refer to a specific panel.

Article Snippet: Antibody , Anti-human CD34 PE (mouse monoclonal) , eBioscience , Clone 4H11 , Flow cytometry 1:200.

Techniques: Gene Expression, Activation Assay, Expressing, Control, Marker, Membrane, Flow Cytometry, Suspension, Derivative Assay

Journal: eLife

Article Title: A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro

doi: 10.7554/eLife.94884

Figure Lengend Snippet: ( A ) Violin plot of IGFBP2 expression profile in the arterial cells obtained from the different conditions, in the presence or absence of gRNAs and doxycycline (DOX). ( B ) Number of haematopoietic colonies obtained after coculture on OP9 in the presence or absence of IGFBP2 (n=3–4 from independent differentiations, **p=0.0080, Sidak’s two-way ANOVA). ( C ) Percentage of DLL4 + arterial cells differentiation within the CD34 + compartment analysed by flow cytometry in day 8 embryoid bodies (EBs) (n=4 from independent differentiations, two-way ANOVA, ns = p>0.99). ( D ) Expansion of haematopoietic progenitors analysed using markers’ expression on suspension progenitors derived after coculture of CD34+ cells onto OP9 support (data are expressed as fold over the CTR in the absence of IGFBP2; n=4 from independent differentiations, *p<0.02, Sidak’s two-way ANOVA). ( E ) Single-cell transcriptomic analysis of developing aorta-gonad-mesonephros (AGM) collected from human embryos at Carnegie stages14 and 15 enriched for CD31+ and CD34+ showing the IGFBP2 expression profile in vivo in the AGM. Figure 4—source data 1. Spreadsheet source file containing the source data used for the plots in . Each tab is labelled to uniquely refer to a specific panel.

Article Snippet: Antibody , Anti-human CD34 PE (mouse monoclonal) , eBioscience , Clone 4H11 , Flow cytometry 1:200.

Techniques: Expressing, Flow Cytometry, Suspension, Derivative Assay, In Vivo

Journal: eLife

Article Title: A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro

doi: 10.7554/eLife.94884

Figure Lengend Snippet: ( A ) Dot plot showing the expression profile of the genes coding for the enzymes mediating glycolysis in CTR and IGFBP2-treated cells at day 10 and 13. ( B ) Dot plot showing the expression profile of the genes coding for the checkpoints of the glycolysis in CTR and IGFBP2-treated cells at days 10 and 13. ( C ) Percentage of cells expressing different markers following OP9 coculture of CD34+ cells (ns for all markers, Kruskall-Wallis one-way ANOVA). ( D ) OP9 cocultures of CD34+ DLL4+ and CD34+DLLL4- (scale bar indicates 50 μm), and in ( E ) their colony formation capacities following 1 week of OP9 coculture (mixed-effect analysis, Sidak’s post-test, ****p<0.0001, **p<0.01). ( F ) Cluster composition in cells treated with IGFBP2 data shows the composition at day 10 and day 13. Figure 5—figure supplement 1—source data 1. Spreadsheet source file containing the source data used for the plots in . Each tab is labelled to uniquely refer to a specific panel.

Article Snippet: Antibody , Anti-human CD34 PE (mouse monoclonal) , eBioscience , Clone 4H11 , Flow cytometry 1:200.

Techniques: Expressing

Journal: eLife

Article Title: A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro

doi: 10.7554/eLife.94884

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-human CD34 PE (mouse monoclonal) , eBioscience , Clone 4H11 , Flow cytometry 1:200.

Techniques: Cloning, Plasmid Preparation, Recombinant, Expressing, Sequencing, Flow Cytometry, Amplification

Journal: eLife

Article Title: A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro

doi: 10.7554/eLife.94884

Figure Lengend Snippet: ( A ) Single-cell transcriptomic analysis of developing aorta-gonad-mesonephros (AGM) collected from human embryos at Carnegie stages 14 and 15 enriched for CD31 + and CD34 + cells from , Nature. Arterial haemogenic endothelium, dashed line, is identified by the colocalisation of DLL4 and RUNX1 . ( B ) Gene expression analysis of the nine selected target genes in the developing human aorta, showing their expression in the haemogenic endothelium.

Article Snippet: Antibody , Anti-human CD34 Percp-Efluor710 (mouse monoclonal) , eBioscience , Clone 4H11 , Flow cytometry 1:100.

Techniques: Gene Expression, Expressing

Journal: eLife

Article Title: A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro

doi: 10.7554/eLife.94884

Figure Lengend Snippet: ( A ) Schematic of the derivation of the inducible SAM (iSAM) cell line by ZNFs mediated targeting of the AAVS1 locus and the subsequent derivation of the iSAM_NT (containing the non-targeting control gRNA) and iSAM_AGM (containing the gRNAs for the target genes) by viral transduction of the gRNAs. ( B ) Schematic of the differentiation protocol with activation of the target genes, used for both the control line iSAM_NT and iSAM_AGM. ( C ) Schematic of the IGFBP2 functional validation experiment. ( D ) Expansion of the arterial population marker by membrane expression of DLL4+ following targets’ activation, quantified by flow cytometry at day 8 of differentiation (*p=0.0190, **p=0.0011, Sidak’s two-way ANOVA). ( E ) Flow cytometry analysis of the cell cycle analysis of suspension progenitor cells was obtained following the OP9 coculture of cells treated with IGFBP2 and control at day 13. ( F ) iSAM expressing cells at day 8 of differentiation upon doxycycline (DOX) treatment. ( G ) Target genes’ expression profile across the different libraries, the colour legend for the libraries is on the top right. ( H ) IGFBP2 expression profile across the cell clusters and in the cell lines and conditions indicated in the title (cluster legend same as in I). ( I ) Contribution of each of the libraries to the cell clusters, grouped by libraries (cluster legend at the bottom). Figure 3—figure supplement 1—source data 1. Spreadsheet source file containing the source data used for the plots in . Each tab is labelled to uniquely refer to a specific panel.

Article Snippet: Antibody , Anti-human CD34 Percp-Efluor710 (mouse monoclonal) , eBioscience , Clone 4H11 , Flow cytometry 1:100.

Techniques: Control, Transduction, Activation Assay, Functional Assay, Biomarker Discovery, Marker, Membrane, Expressing, Flow Cytometry, Cell Cycle Assay, Suspension

Journal: eLife

Article Title: A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro

doi: 10.7554/eLife.94884

Figure Lengend Snippet: ( A ) Gene expression profile of target genes following target genes’ activation, heatmap shows the expression level of the target genes in the iSAM_NT and iSAM_AGM treated with doxycycline (DOX) following normalisation on the -DOX control. ( B ) Dimension reduction and clustering analysis of the scRNAseq data following activation, filtered on cells where the gRNA expression was detected. ( C ) Arterial ( GJA4 , DLL4 ), venous ( NRP2 , APLNR ), and haemogenic marker ( CD44 , RUNX1 ) expression distribution in the clusters indicated by the colour. ( D ) Expression distribution visualised on the UMAP plot showing the location of arterial cells marked by DLL4 , and haemogenic endothelium marked by CD44 and RUNX1 . ( E ) Heatmap of the top 15 marker genes for each of the clusters. ( F ) Contribution of the different libraries to the clusters showing that arterial cell cluster is overrepresented in the iSAM_AGM treated with DOX, compared to the other libraries. ( G ) Expansion of the arterial population assessed by the membrane marker expression of DLL4+ following targets’ activation, quantified by flow cytometry at day 8 of differentiation. (Data are normalised on the iSAM_NT+DOX sample, n=5 independent differentiations, *p=0.0417 paired t-test.) ( H ) Colony-forming potential of the suspension progenitor cells derived from the two lines treated with or without DOX following OP9 coculture activation, data show the colony obtained for 104 CD34+ input equivalent (n=3 from independent differentiations *p<0.05, Tukey’s two-way ANOVA). Figure 3—source data 1. Spreadsheet source file containing the source data used for the plots in . Each tab is labelled to uniquely refer to a specific panel.

Article Snippet: Antibody , Anti-human CD34 Percp-Efluor710 (mouse monoclonal) , eBioscience , Clone 4H11 , Flow cytometry 1:100.

Techniques: Gene Expression, Activation Assay, Expressing, Control, Marker, Membrane, Flow Cytometry, Suspension, Derivative Assay

Journal: eLife

Article Title: A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro

doi: 10.7554/eLife.94884

Figure Lengend Snippet: ( A ) Violin plot of IGFBP2 expression profile in the arterial cells obtained from the different conditions, in the presence or absence of gRNAs and doxycycline (DOX). ( B ) Number of haematopoietic colonies obtained after coculture on OP9 in the presence or absence of IGFBP2 (n=3–4 from independent differentiations, **p=0.0080, Sidak’s two-way ANOVA). ( C ) Percentage of DLL4 + arterial cells differentiation within the CD34 + compartment analysed by flow cytometry in day 8 embryoid bodies (EBs) (n=4 from independent differentiations, two-way ANOVA, ns = p>0.99). ( D ) Expansion of haematopoietic progenitors analysed using markers’ expression on suspension progenitors derived after coculture of CD34+ cells onto OP9 support (data are expressed as fold over the CTR in the absence of IGFBP2; n=4 from independent differentiations, *p<0.02, Sidak’s two-way ANOVA). ( E ) Single-cell transcriptomic analysis of developing aorta-gonad-mesonephros (AGM) collected from human embryos at Carnegie stages14 and 15 enriched for CD31+ and CD34+ showing the IGFBP2 expression profile in vivo in the AGM. Figure 4—source data 1. Spreadsheet source file containing the source data used for the plots in . Each tab is labelled to uniquely refer to a specific panel.

Article Snippet: Antibody , Anti-human CD34 Percp-Efluor710 (mouse monoclonal) , eBioscience , Clone 4H11 , Flow cytometry 1:100.

Techniques: Expressing, Flow Cytometry, Suspension, Derivative Assay, In Vivo

Journal: eLife

Article Title: A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro

doi: 10.7554/eLife.94884

Figure Lengend Snippet: ( A ) Dot plot showing the expression profile of the genes coding for the enzymes mediating glycolysis in CTR and IGFBP2-treated cells at day 10 and 13. ( B ) Dot plot showing the expression profile of the genes coding for the checkpoints of the glycolysis in CTR and IGFBP2-treated cells at days 10 and 13. ( C ) Percentage of cells expressing different markers following OP9 coculture of CD34+ cells (ns for all markers, Kruskall-Wallis one-way ANOVA). ( D ) OP9 cocultures of CD34+ DLL4+ and CD34+DLLL4- (scale bar indicates 50 μm), and in ( E ) their colony formation capacities following 1 week of OP9 coculture (mixed-effect analysis, Sidak’s post-test, ****p<0.0001, **p<0.01). ( F ) Cluster composition in cells treated with IGFBP2 data shows the composition at day 10 and day 13. Figure 5—figure supplement 1—source data 1. Spreadsheet source file containing the source data used for the plots in . Each tab is labelled to uniquely refer to a specific panel.

Article Snippet: Antibody , Anti-human CD34 Percp-Efluor710 (mouse monoclonal) , eBioscience , Clone 4H11 , Flow cytometry 1:100.

Techniques: Expressing

Journal: eLife

Article Title: A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro

doi: 10.7554/eLife.94884

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-human CD34 Percp-Efluor710 (mouse monoclonal) , eBioscience , Clone 4H11 , Flow cytometry 1:100.

Techniques: Cloning, Plasmid Preparation, Recombinant, Expressing, Sequencing, Flow Cytometry, Amplification

Journal: Clinical and Experimental Medicine

Article Title: Mast cell density in Merkel cell carcinoma and its correlation with prognostic features and MCPyV status: a pilot study

doi: 10.1007/s10238-024-01366-4

Figure Lengend Snippet: Vascular density highlighted by CD34 in relation to recurrency/metastasis/death ( a ) and LVI ( b ): note the absence of statistical significance ( p = 0.18 and p = 0.27, respectively)

Article Snippet: All reviewed cases had diagnostic concordance and once the re-evaluation process was completed, new sections were cut by immunostaining with anti-triptase (Monoclonal Mouse, Anti-Human Mast Cell Tryptase, Clone AA1, 1:100 dilution, Dako-Agilent), anti-CD34 (Monoclonal Mouse Anti-Human, Clone QBEnd 10, 1:100 dilution, Dako-Agilent), and anti-MCPyV (large T-antigen, Clone CM2B4, 1:200 dilution, Gennova) antibodies.

Techniques:

Journal: Oncology Letters

Article Title: Peroxiredoxin 2 as a potential prognostic biomarker associated with angiogenesis in cervical squamous cell cancer

doi: 10.3892/ol.2024.14461

Figure Lengend Snippet: Immunohistochemical staining of Prx2, VEGF-A and MVD in CSCC tissues (magnification, ×200). (A) Low expression of (A) Prx2, (B) VEGF-A and (C) MVD (CD34). High expression of (D) Prx2, (E) VEGF-A and (F) MVD (CD34). CSCC, cervical squamous cell cancer; MVD, micro-vessel density; Prx2, peroxiredoxin 2; VEGF-A, vascular endothelial growth factor A.

Article Snippet: The slides were incubated with rabbit polyclonal anti-human Prx2 antibody (Proteintech Group, Inc.; cat. no. 10545-2-AP; 1:500), rabbit polyclonal anti-human VEGF-A antibody (ImmunoWay Biotechnology Company; cat. no. YT5108; 1:100) and mouse monoclonal anti-human CD34 antibody (Dako; Agilent Technologies, Inc.; clone no. QBEnd 10; cat. no. IR632; ready-to-use) overnight at 4°C, followed by incubation at room temperature for 30 min with the Ultra-Sensitive S-P Kit (containing the secondary antibodies; Fuzhou Maixin Biotech Co., Ltd.; cat. no. KIT-9710).

Techniques: Immunohistochemical staining, Staining, Expressing