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R&D Systems cometassay electrophoresis starter kit biotechne r d systems
Cometassay Electrophoresis Starter Kit Biotechne R D Systems, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Figure 1. <t>CRIPTO</t> micro-heterogeneity in ASC population (A) Schematic representation of the experimental design. (B) Representative FACS plots showing the gating strategy. The percentage of CRIPTO-positive cells above the threshold is indicated (bottom panel).

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Presence of ILL-specific CD8 + T cells in the blood of patients with PCa. Cryopreserved PBMCs from HLA-A2 + individuals were cultured for 12 days in the presence <t>of</t> <t>IL-2,</t> IL-15, and either the ILL9mer peptide or the hPAP42mer-mutated peptide. Three groups were compared: healthy individuals, individuals with benign disease, and patients with PCa. On day 12, PBMCs were blocked with a human FcR block and then incubated with surface mAbs and the relevant Dextramer™. ( a ) Proportion of CD8 + T cells within total T cells, ( b ) gating example of Dextramer™-positive CD8 + T cells and proportion of Dextramer™-positive CD8 + T cells following each stimulating condition. ( c ) Cytotoxicity of ILL-stimulated PBMCs against PKH26-labeled WT or PAP low LNCaP cells was measured by flow cytometry. ( d ) Cytotoxicity against PKH26-labeled WT LNCaP cells was compared between each stimulating condition. No difference in the proportion of CD8 + T cells was observed between groups or stimulating conditions. ILL-specific CD8 + T cells were detected following ILL9mer peptide stimulation (benign:1/5, PCa: 6/16) and after hPAP42mer-mutated peptide stimulation (benign: 1/5, PCa: 4/16). Cytotoxicity against LNCaP cells was observed. Bars represent the mean percentage of positive cells and the errors bars represent the SD ( n = 3 to 16 patients per group). Dots represent the mean percentage of cytotoxicity and the errors bars represent the SED ( c ). Differences in the proportion of positive cells between groups were determined using a two-way ANOVA comparison test.

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Figure 1. CRIPTO micro-heterogeneity in ASC population (A) Schematic representation of the experimental design. (B) Representative FACS plots showing the gating strategy. The percentage of CRIPTO-positive cells above the threshold is indicated (bottom panel).

Journal: Developmental cell

Article Title: CRIPTO-based micro-heterogeneity of mouse muscle satellite cells enables adaptive response to regenerative microenvironment.

doi: 10.1016/j.devcel.2023.11.009

Figure Lengend Snippet: Figure 1. CRIPTO micro-heterogeneity in ASC population (A) Schematic representation of the experimental design. (B) Representative FACS plots showing the gating strategy. The percentage of CRIPTO-positive cells above the threshold is indicated (bottom panel).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse Cripto Allophycocyanin MAb Biotechne - R&D systems Cat# FAB1538A Cripto Mouse specific Cell Signaling Technology Cat# 2818; RRID: AB_2240508 Anti-P4HB antibody [RL90] abcam Cat# ab2792; RRID: AB_303304 Pax7 Antibody Developmental Studies Hybridoma Bank (DHSB) Cat# PAX7; RRID: AB_528428 Anti-MyoD (C-20) Santa Cruz Biotechnology, Inc Cat# sc-304; RRID: AB_631992 Anti-GFP antibody abcam Cat# ab13970; RRID: AB_300798 Anti-human Ki67 Monosan Cat# MONX10283 Myogenin Antibody (F5D) Developmental Studies Hybridoma Bank (DHSB) Cat# F5D, RRID: AB_2146602 BrdU antibody [BU1/75 (ICR1)] GeneTex Cat# GTX26326; RRID: AB_1081056 Ki-67 Recombinant Rabbit Monoclonal Antibody (SP6) ThermoFisher Scientific Cat# MA5-14520; RRID: AB_10979488 Anti-Laminin Sigma-Aldrich L9393; RRID: AB_477163 Biotin-SP (long spacer) AffiniPure Goat Anti-Mouse IgG, Fcg subclass 1 specific Jackson ImmunoResearch Cat# 115-065-205, RRID: AB_2338571 Anti-mouse Alexa Fluor 549 ThermoFisher Scientific Cat# A-21203; RRID: AB_2535789 Anti-rabbit Alexa Fluor 488 ThermoFisher Scientific Cat# A-11001; RRID: AB_2534069 Anti-rabbit Alexa Fluor 555 ThermoFisher Scientific Cat# A-31572; RRID: AB_162543 Anti-rabbit Alexa Fluor 549 ThermoFisher Scientific Cat# A-21207; RRID: AB_141637 Alexa Fluor 488 AffiniPure Donkey Anti-Chicken IgY (IgG) (H+L) ThermoFisher Scientific Cat# 703-545-155, RRID: AB_2340375 Cy 3 AffiniPure Donkey Anti-Rat IgG (H+L) Jackson ImmunoResearch Cat# 712-165-153, RRID: AB_2340667 Chemicals, peptides, and recombinant proteins Sambucus Nigra Lectin (SNA, EBL), Biotinylated Vector Laboratories Cat# B-1305-2 Rhodamine Phalloidin ThermoFisher Scientific Cat# R415 Streptavidin-Cy3 conjugated Jackson ImmunoResearch Cat# 016-160-084 Streptavidin-488 conjugated Invitrogen Cat# S32354 Streptavidin-647 conjugated Invitrogen Cat# 532357 Tamoxifene Sigma-Aldrich Cat# T5648 Cardiotoxin Latoxan Laboratory Cat# L8102 Dispase II Roche Cat# 10887800 Collagenase A Roche Cat# 10103586001 DNAse I Roche Cat# 11284932001 Hank’s Balanced Salt Solution (HBSS) Gibco Cat# 14170-088 Bovine Serum Albumin (BSA) Sigma-Aldrich Cat# A4503 Penicillin-Streptomycin Gibco Cat# 15140-122 Collagenase Sigma-Aldrich Cat# C0130 Dulbecco’s Modified Eagle’s medium (DMEM) Gibco Cat# 41965-039 Ham’s F-12 Nutrient Mix, GlutaMAX Gibco Cat# 31765-027 Pyruvate Sodium Gibco Cat# 11360-039 L-Glutamine 200mM (100x) Gibco Cat# 25030-024 (Continued on next page) e1 Developmental Cell 58, 2896–2913.e1–e6, December 18, 2023

Techniques:

Figure 2. Transcriptional proﬁles of CRIPTONeg and CRIPTOPos cells and correlation between CRIPTO and PAX7 expression levels (A) Principal-component analysis (PCA) of CRIPTONeg and CRIPTOPos cells. Each dot represents individual mice. (B) Heatmap of the differential gene expression genes from individual mice (M1–M4). The color scale represents the normalized expression values of each gene.

Journal: Developmental cell

Article Title: CRIPTO-based micro-heterogeneity of mouse muscle satellite cells enables adaptive response to regenerative microenvironment.

doi: 10.1016/j.devcel.2023.11.009

Figure Lengend Snippet: Figure 2. Transcriptional proﬁles of CRIPTONeg and CRIPTOPos cells and correlation between CRIPTO and PAX7 expression levels (A) Principal-component analysis (PCA) of CRIPTONeg and CRIPTOPos cells. Each dot represents individual mice. (B) Heatmap of the differential gene expression genes from individual mice (M1–M4). The color scale represents the normalized expression values of each gene.

Techniques: Expressing, Gene Expression

Figure 4. Regenerative potential of CRIPTONeg and CRIPTOPos cells and reconstitution of naı¨ve CRIPTO micro-heterogeneity following in vivo cell transplantation (A) Schematic representation of the experimental design. (B) Representative images of mosaic recipient muscles showing tdTomato-positive myoﬁbers (RFP-red) and double staining with GFP (green) and LAMININ (white). (C and D) Quantiﬁcation of tdTomato+ myoﬁbers number/area (C) and GFP+ cells from CRIPTONeg and CRIPTOPos donor cells normalized per mm2 (D). Scale bars: 100 mm. Data are mean ± SEM. Nuclei were counter-stained with DAPI. (n = 3). (E) Schematic representation of the experimental design. (F and G) Representative dot plots of surface CRIPTO in GFP-positive CRIPTONeg and CRIPTOPos cells used for transplantation (F) and GFP-positive ASCs (3 dpi) derived from CRIPTONeg and CRIPTOPos -transplanted mice (G). The percentage of CRIPTO-positive cells above the threshold is indicated in the dot plots.

Journal: Developmental cell

Article Title: CRIPTO-based micro-heterogeneity of mouse muscle satellite cells enables adaptive response to regenerative microenvironment.

doi: 10.1016/j.devcel.2023.11.009

Figure Lengend Snippet: Figure 4. Regenerative potential of CRIPTONeg and CRIPTOPos cells and reconstitution of naı¨ve CRIPTO micro-heterogeneity following in vivo cell transplantation (A) Schematic representation of the experimental design. (B) Representative images of mosaic recipient muscles showing tdTomato-positive myoﬁbers (RFP-red) and double staining with GFP (green) and LAMININ (white). (C and D) Quantiﬁcation of tdTomato+ myoﬁbers number/area (C) and GFP+ cells from CRIPTONeg and CRIPTOPos donor cells normalized per mm2 (D). Scale bars: 100 mm. Data are mean ± SEM. Nuclei were counter-stained with DAPI. (n = 3). (E) Schematic representation of the experimental design. (F and G) Representative dot plots of surface CRIPTO in GFP-positive CRIPTONeg and CRIPTOPos cells used for transplantation (F) and GFP-positive ASCs (3 dpi) derived from CRIPTONeg and CRIPTOPos -transplanted mice (G). The percentage of CRIPTO-positive cells above the threshold is indicated in the dot plots.

Techniques: In Vivo, Transplantation Assay, Muscles, Double Staining, Staining, Derivative Assay

Figure 5. Cellular mechanisms of CRIPTO micro-heterogeneity reconstitution (A) Schematic representation of the experimental design. GFP-positive CRIPTONeg and CRIPTOPos ASCs were incubated at RT ± aCRIPTO-APC antibody (line a) or ± U73122 (line b) and stained with aCRIPTO-APC.

Journal: Developmental cell

Article Title: CRIPTO-based micro-heterogeneity of mouse muscle satellite cells enables adaptive response to regenerative microenvironment.

doi: 10.1016/j.devcel.2023.11.009

Figure Lengend Snippet: Figure 5. Cellular mechanisms of CRIPTO micro-heterogeneity reconstitution (A) Schematic representation of the experimental design. GFP-positive CRIPTONeg and CRIPTOPos ASCs were incubated at RT ± aCRIPTO-APC antibody (line a) or ± U73122 (line b) and stained with aCRIPTO-APC.

Techniques: Incubation, Staining

Figure 7. CRIPTO micro-heterogeneity preserves the balance between self-renewal and myogenic commitment (A) Representative pictures of PAX7 (magenta) and GFP (green) staining (upper panel) and quantiﬁcation of PAX7+/GFP± cells (bottom panel) in Criptowt and CriptocKO muscle sections. Inset shows higher magniﬁcation (103) of the boxed area. Scale bars: 75 mm.

Journal: Developmental cell

Article Title: CRIPTO-based micro-heterogeneity of mouse muscle satellite cells enables adaptive response to regenerative microenvironment.

doi: 10.1016/j.devcel.2023.11.009

Figure Lengend Snippet: Figure 7. CRIPTO micro-heterogeneity preserves the balance between self-renewal and myogenic commitment (A) Representative pictures of PAX7 (magenta) and GFP (green) staining (upper panel) and quantiﬁcation of PAX7+/GFP± cells (bottom panel) in Criptowt and CriptocKO muscle sections. Inset shows higher magniﬁcation (103) of the boxed area. Scale bars: 75 mm.

Techniques: Staining

Journal: Cancers

Article Title: A Mutated Prostatic Acid Phosphatase (PAP) Peptide-Based Vaccine Induces PAP-Specific CD8 + T Cells with Ex Vivo Cytotoxic Capacities in HHDII/DR1 Transgenic Mice

doi: 10.3390/cancers14081970

Figure Lengend Snippet: Presence of ILL-specific CD8 + T cells in the blood of patients with PCa. Cryopreserved PBMCs from HLA-A2 + individuals were cultured for 12 days in the presence of IL-2, IL-15, and either the ILL9mer peptide or the hPAP42mer-mutated peptide. Three groups were compared: healthy individuals, individuals with benign disease, and patients with PCa. On day 12, PBMCs were blocked with a human FcR block and then incubated with surface mAbs and the relevant Dextramer™. ( a ) Proportion of CD8 + T cells within total T cells, ( b ) gating example of Dextramer™-positive CD8 + T cells and proportion of Dextramer™-positive CD8 + T cells following each stimulating condition. ( c ) Cytotoxicity of ILL-stimulated PBMCs against PKH26-labeled WT or PAP low LNCaP cells was measured by flow cytometry. ( d ) Cytotoxicity against PKH26-labeled WT LNCaP cells was compared between each stimulating condition. No difference in the proportion of CD8 + T cells was observed between groups or stimulating conditions. ILL-specific CD8 + T cells were detected following ILL9mer peptide stimulation (benign:1/5, PCa: 6/16) and after hPAP42mer-mutated peptide stimulation (benign: 1/5, PCa: 4/16). Cytotoxicity against LNCaP cells was observed. Bars represent the mean percentage of positive cells and the errors bars represent the SD ( n = 3 to 16 patients per group). Dots represent the mean percentage of cytotoxicity and the errors bars represent the SED ( c ). Differences in the proportion of positive cells between groups were determined using a two-way ANOVA comparison test.

Article Snippet: In order to increase the frequency of vaccine-specific CD8 + T cells, splenocytes from each immunized mouse were cultured in 24 well plates (5 × 10 6 /2 mL) for 6 days at 37 °C in T cell medium containing 50 U/mL of recombinant murine IL-2 (R&D Systems, Biotechne, Abingdon, UK), 2 mM of β-mercaptoethanol, and 1 ug/mL ILL 9 mer peptide.

Techniques: Cell Culture, Blocking Assay, Incubation, Labeling, Flow Cytometry, Comparison