Journal: Nature

Article Title: Metformin reduces the competitive advantage of Dnmt3a R878H HSPCs

doi: 10.1038/s41586-025-08871-w

Figure Lengend Snippet: a , Schematic of the metabolic pathways involved in 1C metabolism. B12, vitamin B12; DMG, dimethylglycine; me, methyl; mTHF, methyltetrahydrofolate; THF, tetrahydrofolate. Created in BioRender. Chan, S. (2025) https://BioRender.com/a57k225 . b , Quantification of the indicated metabolites in LK cells isolated from mice transplanted with bone marrow cells of the indicated genotype. The mice were either left untreated or treated with metformin for 1 month. n = 3 biologically independent samples per condition. c , Levels of SAM and SAH and the SAM:SAH ratio in LK cells isolated from mice transplanted with bone marrow cells of the indicated genotype. The mice were either left untreated or treated with metformin for 1 month. n = 12 biologically independent samples for each condition. d , Gene set enrichment plot of bulk RNA-seq data comparing metformin-treated Dnmt3a R878H/+ LK cells ( n = 3 biologically independent samples) versus vehicle-treated Dnmt3a R878H/+ LK cells ( n = 3 biologically independent samples) using the indicated gene set (WP435). ES, enrichment score. e , Expression level of the indicated genes by RT–qPCR in LK cells isolated from mice transplanted with bone marrow cells of the indicated genotype. The mice were either left untreated or treated with metformin for 1 month. n = 3 biologically independent samples for each condition. In b , c , e , the box represents the interquartile range with the median indicated by the line inside the box; whiskers extend to the minimum and maximum values. Statistical significance was calculated using two-sided Student’s t -tests.

Article Snippet: SAM and SAH levels were measured using the SAM and SAH Combo ELISA Kit (Cell Biolabs, STA-671-C).

Techniques: Methylation, Isolation, RNA Sequencing, Expressing, Quantitative RT-PCR

Journal: Nature

Article Title: Metformin reduces the competitive advantage of Dnmt3a R878H HSPCs

doi: 10.1038/s41586-025-08871-w

Figure Lengend Snippet: a , Schematic diagram of the in vitro competition assay of human CD34 + -enriched HSPCs. Created in BioRender. Chan, S. (2025) https://BioRender.com/w95m335 . b , Proportion of BFP + and GFP + cells in a competition assay between BFP + HSPCs expressing shNT, shRNA targeting DNMT3A (sh DNMT3A ) alone or sh DNMT3A plus NDI1 and GFP + HSPCs expressing shNT in the absence or presence of metformin. n = 4 biologically independent samples each. c , Basal OCR in human HSPCs expressing shNT or sh DNMT3A and treated with or without metformin. n = 4 biologically independent samples each. d , Intracellular 5mC staining in human HSPCs expressing the indicated shRNA and treated with or without metformin. n = 3 biologically independent samples each. e , Intracellular H3K27me3 staining in human HSPCs expressing the indicated shRNA and treated with or without metformin. n = 4 biologically independent samples each. f , SAM:SAH ratio in human HSPCs expressing the indicated shRNA and treated with or without metformin. n = 3 biologically independent samples each. g , DNMT3A R882H VAFs of prime-edited human HSPCs at baseline (day 0) and after 14 days in culture in the presence or absence of metformin. n = 5 biologically independent samples. h , Mutant B2M VAFs of prime-edited human HSPCs at baseline (day 0) and after 14 days in culture in the presence or absence of metformin. n = 4 biologically independent samples. Lines connect VAFs from the same sample in g , h . In c – f , the box represents the interquartile range with the median indicated by the line inside the box; whiskers extend to the minimum and maximum values. In b , data are mean ± s.e.m. In g , h , the bar plot represents the mean. Statistical significance was calculated using two-sided unpaired ( b – f ) or paired ( g , h ) Student’s t -tests.

Article Snippet: SAM and SAH levels were measured using the SAM and SAH Combo ELISA Kit (Cell Biolabs, STA-671-C).

Techniques: In Vitro, Competitive Binding Assay, Expressing, shRNA, Staining, Mutagenesis

Journal: Nature

Article Title: Metformin reduces the competitive advantage of Dnmt3a R878H HSPCs

doi: 10.1038/s41586-025-08871-w

Figure Lengend Snippet: a , Expression of DNMT3A mRNA in human CD34 + enriched HSPCs transduced with a lentiviral vector expressing a non-targeting shRNA (shNT) or DNMT3A shRNA. n = 3 technical replicates for 1 biological sample in each condition. b , Intracellular flow cytometry staining for DNMT3A protein in human CD34 + enriched HSPCs transduced with a lentiviral vector expressing a non-targeting shRNA (shNT) or DNMT3A shRNA. The histogram depicting isotype control antibody staining represents cells expressing the non-targeting shRNA (shNT). c , Proportion of BFP + and GFP + cells in a competition assay between BFP + HSPCs expressing shNT or sh DNMT3A and GFP + HSPCs expressing shNT in the absence or presence of metformin at 50 μM. Analysis was gated on the CD34 + cells in the left panel and CD34 - cells in the right panel. n = 2 biologically independent samples for each condition. d , Concentration SAM and SAH in human HSPCs expressing the indicated shRNA and treated with or without metformin. n = 3 biologically independent samples for each condition. e , Representative Sanger sequencing chromatogram of the sequences surrounding the DNMT3A p.R882H (c.2645 G > A) missense mutation in a prime-edited HSPC cell pool. In a, d , the box represents the interquartile range with the median indicated by the line inside the box. Whiskers extend to the minimum and maximum values. In c , bar plots represent mean values. Statistical significance was calculated using two-sided Student’s t-test for all comparisons. *** P < 0.001.

Article Snippet: SAM and SAH levels were measured using the SAM and SAH Combo ELISA Kit (Cell Biolabs, STA-671-C).

Techniques: Expressing, Transduction, Plasmid Preparation, shRNA, Flow Cytometry, Staining, Control, Competitive Binding Assay, Concentration Assay, Sequencing, Mutagenesis

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Posttranslational Modification Defects in Fibroblast Growth Factor Receptor 1 as a Reason for Normosmic Isolated Hypogonadotropic Hypogonadism

doi: 10.1155/2020/2358719

Figure Lengend Snippet: The correlation of genotypes and phenotypes between proband nIHH and reported Kallmann patient.

Article Snippet: Subsequently, the cells were subjected to immunoblotting with anti-p-FGFR1 (1 : 5000, Abcam), anti-FGFR1 (1 : 3500, Abcam), anti-p-ERK (1 : 1000, Cell Signaling Technology), anti-ERK1/2 (1 : 1000, Cell Signaling Technology), anti-p-Akt (1 : 1000, Cell Signaling Technology), anti-Akt (1 : 1000, Cell Signaling Technology), anti-p-STAT3 (1 : 1000, Cell Signaling Technology), anti-STAT3 (1 : 1000, Cell Signaling Technology), and anti-GAPDH (1 : 7000, Proteintech) to observe the expression of the FGFR1 phosphorylation levels as well as the relative phosphorylation levels of the downstream signaling molecules.

Techniques: Mutagenesis

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Posttranslational Modification Defects in Fibroblast Growth Factor Receptor 1 as a Reason for Normosmic Isolated Hypogonadotropic Hypogonadism

doi: 10.1155/2020/2358719

Figure Lengend Snippet: Pedigrees of the proband with FGFR1 and CEP290 mutation. (a) The mutation results in both FGFR1 (A) and CEP290 (B) from all family members. Arrows: mutation sites. (b) Pedigrees of the proband. Circles: females; squares: males; arrows: proband. (c) The domain structures of both FGFR1 (A) and CEP290 (B) with two mutant sites, respectively. SP: signal peptide; HB: binding domain for heparin or heparin sulfate proteoglycan; PLC sites: interaction with PLC gamma; NB: nuclear-binding domain; TK1/2: tyrosine kinase subdomain 1/2; Ig I, Ig II, and Ig III: three Ig-like domains; TM: transmembrane domain.

Article Snippet: Subsequently, the cells were subjected to immunoblotting with anti-p-FGFR1 (1 : 5000, Abcam), anti-FGFR1 (1 : 3500, Abcam), anti-p-ERK (1 : 1000, Cell Signaling Technology), anti-ERK1/2 (1 : 1000, Cell Signaling Technology), anti-p-Akt (1 : 1000, Cell Signaling Technology), anti-Akt (1 : 1000, Cell Signaling Technology), anti-p-STAT3 (1 : 1000, Cell Signaling Technology), anti-STAT3 (1 : 1000, Cell Signaling Technology), and anti-GAPDH (1 : 7000, Proteintech) to observe the expression of the FGFR1 phosphorylation levels as well as the relative phosphorylation levels of the downstream signaling molecules.

Techniques: Mutagenesis, Binding Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Posttranslational Modification Defects in Fibroblast Growth Factor Receptor 1 as a Reason for Normosmic Isolated Hypogonadotropic Hypogonadism

doi: 10.1155/2020/2358719

Figure Lengend Snippet: The bioinformatic assays of the mutations of FGFR1 and CEP290 . (a, b) (A) Conservation analysis of the two mutant sites via multiple sequence alignment. Amino acid in red color are the substitutive amino acid. Asterisks represent a high score of conservation degree. (B) The conservation degree of FGFR1 and CEP290 calculated by WebLogo software. The overall stack height represents the sequence conservation at that position, while the symbol height within the stack indicates the relative frequency of each amino or nucleic acid at that position. (c) The crystal structure of the mutation E670K upon the catalytic (tyrosine kinase) cytosolic domain of FGFR1 and the maps of the coiled-coil domain within mutant and WT CEP290. α -Helices, β -strands, and loops are colored cyan, red, and pink, respectively. Nitrogen and oxygen are colored blue and red, respectively. The mutation sites are labeled with yellow sticks. The phosphorylation site of Tyr-653 and Tyr-638 are shown as green sticks. The activation site is shown as grey sticks. Arrows point to the magnified pictures of selected residues. These structural images are shown using PyMOL.

Article Snippet: Subsequently, the cells were subjected to immunoblotting with anti-p-FGFR1 (1 : 5000, Abcam), anti-FGFR1 (1 : 3500, Abcam), anti-p-ERK (1 : 1000, Cell Signaling Technology), anti-ERK1/2 (1 : 1000, Cell Signaling Technology), anti-p-Akt (1 : 1000, Cell Signaling Technology), anti-Akt (1 : 1000, Cell Signaling Technology), anti-p-STAT3 (1 : 1000, Cell Signaling Technology), anti-STAT3 (1 : 1000, Cell Signaling Technology), and anti-GAPDH (1 : 7000, Proteintech) to observe the expression of the FGFR1 phosphorylation levels as well as the relative phosphorylation levels of the downstream signaling molecules.

Techniques: Mutagenesis, Sequencing, Software, Labeling, Activation Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Posttranslational Modification Defects in Fibroblast Growth Factor Receptor 1 as a Reason for Normosmic Isolated Hypogonadotropic Hypogonadism

doi: 10.1155/2020/2358719

Figure Lengend Snippet: Analysis of subcellular localization and deglycosylation in FGFR1 groups. (a) Subcellular localization of FGFR1 in HEK293 cells. HEK293 cells were transfected with empty vector (EV), FGFR1 (WT), or mutant FGFR1 (p. E670K), and protein localization was observed by fluorescence microscopy. FGFR1 was detected using an anti-FGFR1 antibody followed by secondary antibodies conjugated with Alexa Fluor 555 (red). Nuclei were visualized by DAPI. Original magnification: 600x. (b) Deglycosylation in FGFR1 groups with Endo H and PNGase F. Overall expression levels of FGFR1 in distinct groups were judged from the PNGase F treatments and were normalized to their GAPDH levels, respectively. Maturation analysis was determined from the Endo H-treated groups; the upper band represents the fully glycosylated mature form while the lower band stands for an immature or a core glycosylated form. Percentage of the mature band by density calculations was used to measure the maturation degrees with the groups. Both the results represent the ratio between mutant and WT. EV: empty vector; UT: untreated; E: Endo H-treated; P: PNGase F treated. Arrows point to the molecular weight.

Article Snippet: Subsequently, the cells were subjected to immunoblotting with anti-p-FGFR1 (1 : 5000, Abcam), anti-FGFR1 (1 : 3500, Abcam), anti-p-ERK (1 : 1000, Cell Signaling Technology), anti-ERK1/2 (1 : 1000, Cell Signaling Technology), anti-p-Akt (1 : 1000, Cell Signaling Technology), anti-Akt (1 : 1000, Cell Signaling Technology), anti-p-STAT3 (1 : 1000, Cell Signaling Technology), anti-STAT3 (1 : 1000, Cell Signaling Technology), and anti-GAPDH (1 : 7000, Proteintech) to observe the expression of the FGFR1 phosphorylation levels as well as the relative phosphorylation levels of the downstream signaling molecules.

Techniques: Transfection, Plasmid Preparation, Mutagenesis, Fluorescence, Microscopy, Expressing, Molecular Weight

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Posttranslational Modification Defects in Fibroblast Growth Factor Receptor 1 as a Reason for Normosmic Isolated Hypogonadotropic Hypogonadism

doi: 10.1155/2020/2358719

Figure Lengend Snippet: Functional analysis of FGFR1 in the mutant and WT groups in vitro. (a) Gene expression analysis of FGFR1 in the mutant and WT groups by qPCR. HEK293 cells were transiently transfected with WT, mutant FGFR1 plasmid, or empty vector for RNA extraction, using RT-PCR and real-time quantitative PCR to detect the FGFR1 mRNA expression. (b–d) Gene and cell surface expression analysis of FGFR1 and its downstream signaling. (b) Using RT-PCR and qPCR for the FOS gene expression analysis in the FGF8-induced mutant and WT groups. (c) The phosphorylation levels of WT and mutant FGFR1 and the affected signal pathways tested by western blotting in groups (A). Quantitative analyses of FGFR1 phosphorylation levels (Y653) (B), the total FGFR1 relative level (C), FGF8-induced ERK1/2 (D), and Akt (E) phosphorylation levels are shown with a bar chart. (d) Analysis on the FGFR1-affected JAK/STAT3 pathways by western blotting in groups (A). Quantitative analysis of the STAT3 phosphorylation levels (B). Shown is the meanpercentage ± SD of three biological replicates ( p < 0.01 by Student's t -test).

Article Snippet: Subsequently, the cells were subjected to immunoblotting with anti-p-FGFR1 (1 : 5000, Abcam), anti-FGFR1 (1 : 3500, Abcam), anti-p-ERK (1 : 1000, Cell Signaling Technology), anti-ERK1/2 (1 : 1000, Cell Signaling Technology), anti-p-Akt (1 : 1000, Cell Signaling Technology), anti-Akt (1 : 1000, Cell Signaling Technology), anti-p-STAT3 (1 : 1000, Cell Signaling Technology), anti-STAT3 (1 : 1000, Cell Signaling Technology), and anti-GAPDH (1 : 7000, Proteintech) to observe the expression of the FGFR1 phosphorylation levels as well as the relative phosphorylation levels of the downstream signaling molecules.

Techniques: Functional Assay, Mutagenesis, In Vitro, Expressing, Transfection, Plasmid Preparation, RNA Extraction, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Posttranslational Modification Defects in Fibroblast Growth Factor Receptor 1 as a Reason for Normosmic Isolated Hypogonadotropic Hypogonadism

doi: 10.1155/2020/2358719

Figure Lengend Snippet: A relevance between GnRH-deficiency-related signaling network and several known disease-causing genes of olfactory dysfunction by using STRING software. The protein-protein interaction network encoded by pathogenic genes known to cause KS including FGFR1 (colored blue) were correlated with CEP290-related protein-protein network involving olfactory dysfunction via several pathways containing the factors of FGF8, NTRK2, AHI1, and IFT88.

Article Snippet: Subsequently, the cells were subjected to immunoblotting with anti-p-FGFR1 (1 : 5000, Abcam), anti-FGFR1 (1 : 3500, Abcam), anti-p-ERK (1 : 1000, Cell Signaling Technology), anti-ERK1/2 (1 : 1000, Cell Signaling Technology), anti-p-Akt (1 : 1000, Cell Signaling Technology), anti-Akt (1 : 1000, Cell Signaling Technology), anti-p-STAT3 (1 : 1000, Cell Signaling Technology), anti-STAT3 (1 : 1000, Cell Signaling Technology), and anti-GAPDH (1 : 7000, Proteintech) to observe the expression of the FGFR1 phosphorylation levels as well as the relative phosphorylation levels of the downstream signaling molecules.

Techniques: Software

Journal: Molecular Cancer

Article Title: Proteinase-activated receptor 2 (PAR 2 ) in hepatic stellate cells – evidence for a role in hepatocellular carcinoma growth in vivo

doi: 10.1186/s12943-016-0538-y

Figure Lengend Snippet: PAR 2 knockdown in LX-2 cells inhibits tumour growth in a SCID mouse model. a - c Expression and function of PAR 2 in LX-2 cells. a RT-PCR of PAR 2 expression. Extraction of total RNA from the LX-2-wt cells and synthesis of cDNA was performed as described in the section. PCR reactions without cDNAs were run as a negative control (Primer). Integrity of the cDNA was independently confirmed by amplification of beta-actin (Actin). MW marker, molecular-weight marker. Representative results of three independent experiments are shown. b PAR 2 immunofluorescence was detected using the confocal laser scanning microscope LSM-510 Meta (Carl Zeiss, Germany). Localization of immunofluorescence labelled PAR 2 is shown in permeabilized LX-2-wt cells using SAM-11 (1:100) and a FITC-conjugated anti-mouse IgG (1:200) as secondary antibody. c LX-2-wt cells grown on Lab Tek chambered borosilicate cover glass were loaded with fluo-4-AM as described in . For calcium measurements, an inverted confocal laser scanning microscope LSM 510 was used. Fluorescence was monitored at 488 nm. (a) PAR 2 -AP (10 µM) and (b) trypsin (10 nM) induce Ca 2+ rise in LX-2 cells. (c) Fluorescence images, in pseudocolor, from single LX-2 cells. The sequence shows a fast and transient fluorescence increase from 15 s to 40 s after PAR 2 -AP addition (0 s). Data represent the mean ± SD from calcium measurements in 20 individual cells, respectively. ( d - f ) PAR 2 knockdown in LX-2 cells inhibits tumour growth in a SCID mouse model. SCID mice were randomized into five groups, each consisting of 8 animals. Hep3B and LX-2 cells were subcutaneously (co)injected at the right flank of the mice [(1): 5 × 10 5 LX-2-wt; (2): 10 5 Hep3B cells; (3): 10 5 Hep3B cells plus 5 × 10 5 LX-2-wt, (4): 10 5 Hep3B cells plus 5 × 10 5 LX-2-shCo cells, (5): 10 5 Hep3B cells plus 5 × 10 5 LX-2-shPAR 2 cells]. d Tumours prepared 16 days after xenotransplantation. Data for body weight and tumour volume are indicated as mean ± SD ( n = 8/group); ** p < 0.01). e Representative micro CT images from the tumours 16 days after xenotransplantation. For 3D reconstruction of the tumours the software “Imalytics” (Philips GmbH, Aachen, Germany) was used. f Paraffin sections obtained from tumours and stained for hematoxylin/eosin, cytokeratin, Ki67 or CD31. Images were taken at magnification = × 5 ( left panel ) and at magnification = × 200

Article Snippet: The expression of PAR 2 on protein level was established using the monoclonal anti-PAR 2 antibody SAM-11, generated against a peptide representing amino acids 37 SLIGKVDGTSHVTG 50 of human PAR 2 (Santa Cruz Biotechnology).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Negative Control, Amplification, Marker, Molecular Weight, Immunofluorescence, Laser-Scanning Microscopy, Fluorescence, Sequencing, Injection, Micro-CT, Software, Staining

Journal: Molecular Cancer

Article Title: Proteinase-activated receptor 2 (PAR 2 ) in hepatic stellate cells – evidence for a role in hepatocellular carcinoma growth in vivo

doi: 10.1186/s12943-016-0538-y

Figure Lengend Snippet: PAR 2 knockdown inhibits the pro-mitotic effect of LX-2 cell culture supernatants on Hep3B cells. LX-2 cells were treated on two consecutive days with transfection agent (Lipofectamine 2000) alone (-) or Lipofectamine 2000 plus 50 nM each of a universal control siRNA (Co) or siRNA to PAR 1 or PAR 2 . After the second transfection, cells received normal growth medium which remained on the cells for 48 h. The resulting culture supernatants from PAR 2 or PAR 1 depleted and control LX-2 cells were then harvested, cleared by centrifugation and transferred to cultures of Hep3B cells. After a 72 h incubation period with the LX-2 conditioned media, Hep3B cells were detached and cell numbers determined by manual counting and trypan blue exclusion test. Data represent relative cell numbers (mean ± S.D.) from three independent experiments with cell numbers of Co cells set at 100 %. * p < 0.05 versus control

Article Snippet: The expression of PAR 2 on protein level was established using the monoclonal anti-PAR 2 antibody SAM-11, generated against a peptide representing amino acids 37 SLIGKVDGTSHVTG 50 of human PAR 2 (Santa Cruz Biotechnology).

Techniques: Cell Culture, Transfection, Centrifugation, Incubation

Journal: Molecular Cancer

Article Title: Proteinase-activated receptor 2 (PAR 2 ) in hepatic stellate cells – evidence for a role in hepatocellular carcinoma growth in vivo

doi: 10.1186/s12943-016-0538-y

Figure Lengend Snippet: LX-2 cells stimulated with PAR 2 -AP secrete enhanced level of proliferation- and angiogenesis-associated cytokines and proteinases. LX-2-wt cells, serum-starved for 17 h were treated with PAR 2 -AP (10 μM), trypsin (10 nM) or vehicle for 24 h and the supernatants were evaluated by protein analyses using a Human Angiogenesis Array Kit and b Human Protease Array kit as described in the Method section. In the representative images each pair of horizontal blots (bands) represents a different protein present in the supernatant, whereas the intensities of the blots characterised the amount of the respective protein (R = reference spot for protein loading). The results in the histograms below the images represent the mean ± SD for three independent experiments [**indicates a significant difference from non-stimulated ( p < 0.05)]

Article Snippet: The expression of PAR 2 on protein level was established using the monoclonal anti-PAR 2 antibody SAM-11, generated against a peptide representing amino acids 37 SLIGKVDGTSHVTG 50 of human PAR 2 (Santa Cruz Biotechnology).

Techniques:

Journal: Molecular Cancer

Article Title: Proteinase-activated receptor 2 (PAR 2 ) in hepatic stellate cells – evidence for a role in hepatocellular carcinoma growth in vivo

doi: 10.1186/s12943-016-0538-y

Figure Lengend Snippet: PAR 2 activation mediates enhanced migration of LX-2 cells in a Met-, PDGFR-, Src- kinase-, p42/p44 MAPK- and MMP-dependent manner. a LX-2 cells (wt, shPAR 2 or shCo as indicated) were serum-starved for 17 h and treated with PAR 2 -AP (10 μM), or trypsin (10 nM) for 6 h. Cells had migrated through the collagen barrier and the pores of the polycarbonate membrane were fixed, stained and quantified by microscopic counting. Bars represent the mean values ± S.D. of octuplicates obtained in one experiment, which is representative for three independent assays. ** P -value < 0.05 versus non stimulated control. b Serum-starved LX-2-wt cells were stimulated with PAR 2 -AP (10 μM) for 4 h and analysis of plasma membrane filopodial structures was performed using scanning electron microscopy as described in the method section. ( a ) Cells with filopodia were quantified in a blinded manner from 100 individual cells per each group and from three experimental preparations. ( b ) enlarged inset. The picture shows a single LX-2-wt cell with typical filopodial spike pattern after stimulation with PAR 2 -AP. c Serum-starved LX-2-wt cells were preincubated for 1 h with vehicle, SU 11274 (10 μM), PHA 665752 (0.1 μM), AG 1296 (1.0 μM), SL 372 (5.0 μM), PD 98059 (10 μM), U 0126 (10 μM), PP2 (5.0 μM), or GM-6001 (1.0 μM). Cell migration in response to PAR 2 -AP (10 μM) was analysed after 6 h as described under a . Representative results from three independent experiments are shown. * P -value < 0.05 versus non stimulated control, ** P -value < 0.05 versus stimulated with PAR 2 -AP and versus non stimulated control

Article Snippet: The expression of PAR 2 on protein level was established using the monoclonal anti-PAR 2 antibody SAM-11, generated against a peptide representing amino acids 37 SLIGKVDGTSHVTG 50 of human PAR 2 (Santa Cruz Biotechnology).

Techniques: Activation Assay, Migration, Staining, Electron Microscopy