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p ddr1 2  (R&D Systems)


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    R&D Systems p ddr1 2
    MAPK inhibitor-resistant melanoma cells exhibit <t>enhanced</t> <t>DDR1/2</t> signaling. ( A ) Clonogenic assay showing the validation of drug-resistant cell lines. ( B ) Quantification of clonogenic assay shown in Fig. 1A. Data are presented as relative colony formation (%) normalized to the control group and expressed as the mean ± SEM, n = 3 replicates. Statistical significance was determined by two-way ANOVA. * p < 0.05, # p < 0.001, † p < 0.001, ‡ p < 0.0001. ( C ) Cell viability assay of RAF and MEK inhibitors in parental cells and drug-resistant cells for 48 h. Data are presented as the mean ± SEM, n = 3 replicates. ( D ) Western blot analysis of pERK and pAKT signaling responses in parental and resistant melanoma cells following 48-h treatment with vehicle or the corresponding resistance-selecting drug(s) at 1 µM. ( E ) Bubble plot representing Gene Set Enrichment Analysis (GSEA) using hallmark gene set comparing parental and resistant melanoma cells. Pathway enrichment was calculated using GSEA, with color indicating NES (blue = downregulated, red = upregulated) and bubble size representing −log10(p-value). ( F ) SK-MEL-2 and SK-MEL-2BR cells were stimulated with type I collagen (40 µg/ml) for 18 h to activate DDR signaling
    P Ddr1 2, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/ddr2/Human+Phospho-DDR1%2FDDR2+(DDR1+Y796%2C+DDR2+Y740)+Antibody/pmc13063777-64-28-29
    Average 93 stars, based on 30 article reviews
    p ddr1 2 - by Bioz Stars, 2026-09
    93/100 stars

    Images

    1) Product Images from "PHI-501, a dual inhibitor of RAF and DDR1/2, overcomes MAPK drug resistance in Melanoma"

    Article Title: PHI-501, a dual inhibitor of RAF and DDR1/2, overcomes MAPK drug resistance in Melanoma

    Journal: Cancer Cell International

    doi: 10.1186/s12935-026-04271-w

    MAPK inhibitor-resistant melanoma cells exhibit enhanced DDR1/2 signaling. ( A ) Clonogenic assay showing the validation of drug-resistant cell lines. ( B ) Quantification of clonogenic assay shown in Fig. 1A. Data are presented as relative colony formation (%) normalized to the control group and expressed as the mean ± SEM, n = 3 replicates. Statistical significance was determined by two-way ANOVA. * p < 0.05, # p < 0.001, † p < 0.001, ‡ p < 0.0001. ( C ) Cell viability assay of RAF and MEK inhibitors in parental cells and drug-resistant cells for 48 h. Data are presented as the mean ± SEM, n = 3 replicates. ( D ) Western blot analysis of pERK and pAKT signaling responses in parental and resistant melanoma cells following 48-h treatment with vehicle or the corresponding resistance-selecting drug(s) at 1 µM. ( E ) Bubble plot representing Gene Set Enrichment Analysis (GSEA) using hallmark gene set comparing parental and resistant melanoma cells. Pathway enrichment was calculated using GSEA, with color indicating NES (blue = downregulated, red = upregulated) and bubble size representing −log10(p-value). ( F ) SK-MEL-2 and SK-MEL-2BR cells were stimulated with type I collagen (40 µg/ml) for 18 h to activate DDR signaling
    Figure Legend Snippet: MAPK inhibitor-resistant melanoma cells exhibit enhanced DDR1/2 signaling. ( A ) Clonogenic assay showing the validation of drug-resistant cell lines. ( B ) Quantification of clonogenic assay shown in Fig. 1A. Data are presented as relative colony formation (%) normalized to the control group and expressed as the mean ± SEM, n = 3 replicates. Statistical significance was determined by two-way ANOVA. * p < 0.05, # p < 0.001, † p < 0.001, ‡ p < 0.0001. ( C ) Cell viability assay of RAF and MEK inhibitors in parental cells and drug-resistant cells for 48 h. Data are presented as the mean ± SEM, n = 3 replicates. ( D ) Western blot analysis of pERK and pAKT signaling responses in parental and resistant melanoma cells following 48-h treatment with vehicle or the corresponding resistance-selecting drug(s) at 1 µM. ( E ) Bubble plot representing Gene Set Enrichment Analysis (GSEA) using hallmark gene set comparing parental and resistant melanoma cells. Pathway enrichment was calculated using GSEA, with color indicating NES (blue = downregulated, red = upregulated) and bubble size representing −log10(p-value). ( F ) SK-MEL-2 and SK-MEL-2BR cells were stimulated with type I collagen (40 µg/ml) for 18 h to activate DDR signaling

    Techniques Used: Clonogenic Assay, Biomarker Discovery, Control, Viability Assay, Western Blot

    DDR1 and DDR2 are associated with MAPK pathway activation and therapeutic resistance in melanoma. ( A ) Kaplan–Meier survival analysis showing patient outcomes based on DDR1 and DDR2 expression levels across all TCGA cancer types. Patients were stratified into high (top 25%) and low (bottom 25%) expression groups, and overall survival (OS) was analyzed. ( B ) Proteomic analysis of BRAF/NRAS-mutant SKCM. Box plots demonstrate differential expression of AKT and phosphorylated MEK1 (pMEK1-S217/S221) between wild-type and mutant groups. ( C ) Correlation heatmaps between DDR1/2 and key genes in the MAPK and AKT signaling pathways in TCGA-SKCM samples. Color intensity represents the correlation value, with red indicating a positive correlation. Significance levels are indicated by asterisks (* p < 0.05, ** p < 0.01, *** p < 0.001). ( D ) Correlation analysis revealing DDR1 expression (DepMap) and MAPK pathway inhibitor sensitivity (GDSC2) in melanoma cell lines. Scatter plots depicting correlations between DDR1 expression levels and IC50 to Dabrafenib (left) and Trametinib (right)
    Figure Legend Snippet: DDR1 and DDR2 are associated with MAPK pathway activation and therapeutic resistance in melanoma. ( A ) Kaplan–Meier survival analysis showing patient outcomes based on DDR1 and DDR2 expression levels across all TCGA cancer types. Patients were stratified into high (top 25%) and low (bottom 25%) expression groups, and overall survival (OS) was analyzed. ( B ) Proteomic analysis of BRAF/NRAS-mutant SKCM. Box plots demonstrate differential expression of AKT and phosphorylated MEK1 (pMEK1-S217/S221) between wild-type and mutant groups. ( C ) Correlation heatmaps between DDR1/2 and key genes in the MAPK and AKT signaling pathways in TCGA-SKCM samples. Color intensity represents the correlation value, with red indicating a positive correlation. Significance levels are indicated by asterisks (* p < 0.05, ** p < 0.01, *** p < 0.001). ( D ) Correlation analysis revealing DDR1 expression (DepMap) and MAPK pathway inhibitor sensitivity (GDSC2) in melanoma cell lines. Scatter plots depicting correlations between DDR1 expression levels and IC50 to Dabrafenib (left) and Trametinib (right)

    Techniques Used: Activation Assay, Expressing, Mutagenesis, Quantitative Proteomics, Protein-Protein interactions

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    Control:

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart.
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: TRPV4 regulates β1 integrin-mediated cell-matrix adhesions and collagen remodeling.
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    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
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    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
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    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
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    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).

    Expressing:

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart.
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: TRPV4 regulates β1 integrin-mediated cell-matrix adhesions and collagen remodeling.
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    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).. Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to the protocol provided. siRNA transfection Cells with > 80% sub-confluency were transfected with β-Catenin siRNA using Lipofectamine RNAiMAX (Invitrogen).Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).

    Derivative Assay:

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart.
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: TRPV4 regulates β1 integrin-mediated cell-matrix adhesions and collagen remodeling.
    Article Snippet: GSK1016790A (a TRPV4 agonist), ionomycin (Ca2+ ionophore), bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma.bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma. ... The source of information for primary antibodies used is as follows: TRPV4 (ab39260 and ab191580, Abcam; MABS466, Millipore); β1- integrin (ab1952 and MAB1997, Millipore; clone KMI6, 14- 0292- 82, Invitrogen; clone 9EG7, 553 715, BD Biosciences); β3integrin (13166S, Cell Signaling); β5- integrin (3629S, Cell Signaling); α2- integrin (ab181548, Abcam); α11- integrin (ab198826, Abcam); α5- integrin (sc10729, Santa Cruz Biotechnology); αV- integrin (BD611012, BD Biosciences); β- actin (A1978, Millipore); DDR1 (sc- 374618, Santa Cruz Biotechnology); DDR2 (AF2538, R&D systems); Na+/K+ATPase α1 (3010S, Cell Signaling); GAPDH (MAB374, Millipore); Paxillin (ab32084, Abcam); Vimentin (NB300- 223, Novus Biologicals); Collagen I (PA5- 95137, Invitrogen); GFP (A- 11122, Invitrogen).. IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from LI- COR: Goat anti- Mouse (926- 68070; 926- 32210); Goat anti- Rabbit (926- 68071; 926- 32211); Donkey anti- Goat (926- 32214).IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).. Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to the protocol provided. siRNA transfection Cells with > 80% sub-confluency were transfected with β-Catenin siRNA using Lipofectamine RNAiMAX (Invitrogen).Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).

    Immunostaining:

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart.
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: TRPV4 regulates β1 integrin-mediated cell-matrix adhesions and collagen remodeling.
    Article Snippet: GSK1016790A (a TRPV4 agonist), ionomycin (Ca2+ ionophore), bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma.bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma. ... The source of information for primary antibodies used is as follows: TRPV4 (ab39260 and ab191580, Abcam; MABS466, Millipore); β1- integrin (ab1952 and MAB1997, Millipore; clone KMI6, 14- 0292- 82, Invitrogen; clone 9EG7, 553 715, BD Biosciences); β3integrin (13166S, Cell Signaling); β5- integrin (3629S, Cell Signaling); α2- integrin (ab181548, Abcam); α11- integrin (ab198826, Abcam); α5- integrin (sc10729, Santa Cruz Biotechnology); αV- integrin (BD611012, BD Biosciences); β- actin (A1978, Millipore); DDR1 (sc- 374618, Santa Cruz Biotechnology); DDR2 (AF2538, R&D systems); Na+/K+ATPase α1 (3010S, Cell Signaling); GAPDH (MAB374, Millipore); Paxillin (ab32084, Abcam); Vimentin (NB300- 223, Novus Biologicals); Collagen I (PA5- 95137, Invitrogen); GFP (A- 11122, Invitrogen).. IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from LI- COR: Goat anti- Mouse (926- 68070; 926- 32210); Goat anti- Rabbit (926- 68071; 926- 32211); Donkey anti- Goat (926- 32214).IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).. Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to the protocol provided. siRNA transfection Cells with > 80% sub-confluency were transfected with β-Catenin siRNA using Lipofectamine RNAiMAX (Invitrogen).Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).

    Marker:

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart.
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: TRPV4 regulates β1 integrin-mediated cell-matrix adhesions and collagen remodeling.
    Article Snippet: GSK1016790A (a TRPV4 agonist), ionomycin (Ca2+ ionophore), bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma.bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma. ... The source of information for primary antibodies used is as follows: TRPV4 (ab39260 and ab191580, Abcam; MABS466, Millipore); β1- integrin (ab1952 and MAB1997, Millipore; clone KMI6, 14- 0292- 82, Invitrogen; clone 9EG7, 553 715, BD Biosciences); β3integrin (13166S, Cell Signaling); β5- integrin (3629S, Cell Signaling); α2- integrin (ab181548, Abcam); α11- integrin (ab198826, Abcam); α5- integrin (sc10729, Santa Cruz Biotechnology); αV- integrin (BD611012, BD Biosciences); β- actin (A1978, Millipore); DDR1 (sc- 374618, Santa Cruz Biotechnology); DDR2 (AF2538, R&D systems); Na+/K+ATPase α1 (3010S, Cell Signaling); GAPDH (MAB374, Millipore); Paxillin (ab32084, Abcam); Vimentin (NB300- 223, Novus Biologicals); Collagen I (PA5- 95137, Invitrogen); GFP (A- 11122, Invitrogen).. IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from LI- COR: Goat anti- Mouse (926- 68070; 926- 32210); Goat anti- Rabbit (926- 68071; 926- 32211); Donkey anti- Goat (926- 32214).IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).. Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to the protocol provided. siRNA transfection Cells with > 80% sub-confluency were transfected with β-Catenin siRNA using Lipofectamine RNAiMAX (Invitrogen).Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).

    Staining:

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart.
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: TRPV4 regulates β1 integrin-mediated cell-matrix adhesions and collagen remodeling.
    Article Snippet: GSK1016790A (a TRPV4 agonist), ionomycin (Ca2+ ionophore), bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma.bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma. ... The source of information for primary antibodies used is as follows: TRPV4 (ab39260 and ab191580, Abcam; MABS466, Millipore); β1- integrin (ab1952 and MAB1997, Millipore; clone KMI6, 14- 0292- 82, Invitrogen; clone 9EG7, 553 715, BD Biosciences); β3integrin (13166S, Cell Signaling); β5- integrin (3629S, Cell Signaling); α2- integrin (ab181548, Abcam); α11- integrin (ab198826, Abcam); α5- integrin (sc10729, Santa Cruz Biotechnology); αV- integrin (BD611012, BD Biosciences); β- actin (A1978, Millipore); DDR1 (sc- 374618, Santa Cruz Biotechnology); DDR2 (AF2538, R&D systems); Na+/K+ATPase α1 (3010S, Cell Signaling); GAPDH (MAB374, Millipore); Paxillin (ab32084, Abcam); Vimentin (NB300- 223, Novus Biologicals); Collagen I (PA5- 95137, Invitrogen); GFP (A- 11122, Invitrogen).. IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from LI- COR: Goat anti- Mouse (926- 68070; 926- 32210); Goat anti- Rabbit (926- 68071; 926- 32211); Donkey anti- Goat (926- 32214).IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).. Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to the protocol provided. siRNA transfection Cells with > 80% sub-confluency were transfected with β-Catenin siRNA using Lipofectamine RNAiMAX (Invitrogen).Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).

    Irradiation:

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart.
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: TRPV4 regulates β1 integrin-mediated cell-matrix adhesions and collagen remodeling.
    Article Snippet: GSK1016790A (a TRPV4 agonist), ionomycin (Ca2+ ionophore), bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma.bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma. ... The source of information for primary antibodies used is as follows: TRPV4 (ab39260 and ab191580, Abcam; MABS466, Millipore); β1- integrin (ab1952 and MAB1997, Millipore; clone KMI6, 14- 0292- 82, Invitrogen; clone 9EG7, 553 715, BD Biosciences); β3integrin (13166S, Cell Signaling); β5- integrin (3629S, Cell Signaling); α2- integrin (ab181548, Abcam); α11- integrin (ab198826, Abcam); α5- integrin (sc10729, Santa Cruz Biotechnology); αV- integrin (BD611012, BD Biosciences); β- actin (A1978, Millipore); DDR1 (sc- 374618, Santa Cruz Biotechnology); DDR2 (AF2538, R&D systems); Na+/K+ATPase α1 (3010S, Cell Signaling); GAPDH (MAB374, Millipore); Paxillin (ab32084, Abcam); Vimentin (NB300- 223, Novus Biologicals); Collagen I (PA5- 95137, Invitrogen); GFP (A- 11122, Invitrogen).. IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from LI- COR: Goat anti- Mouse (926- 68070; 926- 32210); Goat anti- Rabbit (926- 68071; 926- 32211); Donkey anti- Goat (926- 32214).IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).. Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to the protocol provided. siRNA transfection Cells with > 80% sub-confluency were transfected with β-Catenin siRNA using Lipofectamine RNAiMAX (Invitrogen).Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).

    Activation Assay:

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart.
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: TRPV4 regulates β1 integrin-mediated cell-matrix adhesions and collagen remodeling.
    Article Snippet: GSK1016790A (a TRPV4 agonist), ionomycin (Ca2+ ionophore), bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma.bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma. ... The source of information for primary antibodies used is as follows: TRPV4 (ab39260 and ab191580, Abcam; MABS466, Millipore); β1- integrin (ab1952 and MAB1997, Millipore; clone KMI6, 14- 0292- 82, Invitrogen; clone 9EG7, 553 715, BD Biosciences); β3integrin (13166S, Cell Signaling); β5- integrin (3629S, Cell Signaling); α2- integrin (ab181548, Abcam); α11- integrin (ab198826, Abcam); α5- integrin (sc10729, Santa Cruz Biotechnology); αV- integrin (BD611012, BD Biosciences); β- actin (A1978, Millipore); DDR1 (sc- 374618, Santa Cruz Biotechnology); DDR2 (AF2538, R&D systems); Na+/K+ATPase α1 (3010S, Cell Signaling); GAPDH (MAB374, Millipore); Paxillin (ab32084, Abcam); Vimentin (NB300- 223, Novus Biologicals); Collagen I (PA5- 95137, Invitrogen); GFP (A- 11122, Invitrogen).. IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from LI- COR: Goat anti- Mouse (926- 68070; 926- 32210); Goat anti- Rabbit (926- 68071; 926- 32211); Donkey anti- Goat (926- 32214).IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).. Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to the protocol provided. siRNA transfection Cells with > 80% sub-confluency were transfected with β-Catenin siRNA using Lipofectamine RNAiMAX (Invitrogen).Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).

    Immunofluorescence:

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart.
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: TRPV4 regulates β1 integrin-mediated cell-matrix adhesions and collagen remodeling.
    Article Snippet: GSK1016790A (a TRPV4 agonist), ionomycin (Ca2+ ionophore), bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma.bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma. ... The source of information for primary antibodies used is as follows: TRPV4 (ab39260 and ab191580, Abcam; MABS466, Millipore); β1- integrin (ab1952 and MAB1997, Millipore; clone KMI6, 14- 0292- 82, Invitrogen; clone 9EG7, 553 715, BD Biosciences); β3integrin (13166S, Cell Signaling); β5- integrin (3629S, Cell Signaling); α2- integrin (ab181548, Abcam); α11- integrin (ab198826, Abcam); α5- integrin (sc10729, Santa Cruz Biotechnology); αV- integrin (BD611012, BD Biosciences); β- actin (A1978, Millipore); DDR1 (sc- 374618, Santa Cruz Biotechnology); DDR2 (AF2538, R&D systems); Na+/K+ATPase α1 (3010S, Cell Signaling); GAPDH (MAB374, Millipore); Paxillin (ab32084, Abcam); Vimentin (NB300- 223, Novus Biologicals); Collagen I (PA5- 95137, Invitrogen); GFP (A- 11122, Invitrogen).. IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from LI- COR: Goat anti- Mouse (926- 68070; 926- 32210); Goat anti- Rabbit (926- 68071; 926- 32211); Donkey anti- Goat (926- 32214).IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).. Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to the protocol provided. siRNA transfection Cells with > 80% sub-confluency were transfected with β-Catenin siRNA using Lipofectamine RNAiMAX (Invitrogen).Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).

    Transplantation Assay:

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart.
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: TRPV4 regulates β1 integrin-mediated cell-matrix adhesions and collagen remodeling.
    Article Snippet: GSK1016790A (a TRPV4 agonist), ionomycin (Ca2+ ionophore), bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma.bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma. ... The source of information for primary antibodies used is as follows: TRPV4 (ab39260 and ab191580, Abcam; MABS466, Millipore); β1- integrin (ab1952 and MAB1997, Millipore; clone KMI6, 14- 0292- 82, Invitrogen; clone 9EG7, 553 715, BD Biosciences); β3integrin (13166S, Cell Signaling); β5- integrin (3629S, Cell Signaling); α2- integrin (ab181548, Abcam); α11- integrin (ab198826, Abcam); α5- integrin (sc10729, Santa Cruz Biotechnology); αV- integrin (BD611012, BD Biosciences); β- actin (A1978, Millipore); DDR1 (sc- 374618, Santa Cruz Biotechnology); DDR2 (AF2538, R&D systems); Na+/K+ATPase α1 (3010S, Cell Signaling); GAPDH (MAB374, Millipore); Paxillin (ab32084, Abcam); Vimentin (NB300- 223, Novus Biologicals); Collagen I (PA5- 95137, Invitrogen); GFP (A- 11122, Invitrogen).. IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from LI- COR: Goat anti- Mouse (926- 68070; 926- 32210); Goat anti- Rabbit (926- 68071; 926- 32211); Donkey anti- Goat (926- 32214).IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).. Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to the protocol provided. siRNA transfection Cells with > 80% sub-confluency were transfected with β-Catenin siRNA using Lipofectamine RNAiMAX (Invitrogen).Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart.
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: Fibroblasts facilitate lymphatic vessel formation in transplanted heart
    Article Snippet: Cryosections were aired-dried for 30 minutes at room temperature, blocked with 5% donkey serum (Solarbio®, SL050) containing 0.1% Triton X-100(Sigma, X-100) in PBS at room temperature for 1 h, stained sections with primary antibodies overnight at 4 °C, and then incubated with or without Alexa Fluor-conjugated secondary antibodies (Invitrogen, 1:500) at room temperature for 1 h. Then, DAPI (Servicebio, G1012) was stained for 15 min at room temperature before being mounted with an anti-fade mounting medium (Servicebio, G1401).. Primary antibodies were applied in this study as listed: LYVE1 (abcam, ab14917, 1:300), VEGFR3 (R&D Systems, AF349, 1:50), tdTomato (Rockland, 600-401-379, 1:500), Periostin (R&D system, AF2955, 1:50), Collagen Type I (Proteintech, 67288-1-Ig, 1:100 ), DDR2 (discoidin domain receptor 2, R&D Systems, MAB25381, 1:100), PDGFRα (R&D Systems, AF1062, 1:50), PCNA (Abclonal, A0264, 1:500), KI67 (abcam, ab16667, 1:500), VEGFD (Proteintech, 26915-1-AP, 1:50), Midkine (Proteintech, 11009-1-AP, 1:50), SEMA3C (R&D Systems,MAB1728, 1:50), PTN (abcam, ab79411, 1:50), NRP2 (Neuropilin-2, CST, D39A5, 1:50), NCL (nucleolin, Santa cruz, sc-8031, 1:50), Vimentin (abcam, ab8978, 1:100), Fibronectin (abcam, ab2413, 1:50), CD31 (R&D, AF3628, 1:50), CD31 (abcam, ab28364, 1:50), SMA-FITC (Sigma, F3777, 1:300), p-AKT(Santa cruz, sc-514032, 1:50), p-ERK (Santa cruz, sc-7383, 1:50), CD45 (R&D Systems, AF114, 1:50), TNF-α(Proteintech, 60291-1-Ig, 1:50), IL1β (Abclonal, a19635, 1:50), IL6 (Proteintech, 66146-1-Ig, 1:50).. Secondary antibodies were used as listed: donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen, A-21206, 1:500), donkey anti-rabbit IgG Alexa Fluor 555 (Invitrogen, A-31572, 1:500), donkey anti-rat IgG Alexa Fluor 488 (Invitrogen, A-21208, 1:500), donkey anti-mouse IgG Alexa Fluor 488 (Invitrogen, A-21202, 1:500), donkey anti-mouse IgG Alexa Fluor 555(Invitrogen, A-31570, 1:500), donkey anti-mouse IgG Alexa Fluor 647(Invitrogen, A-32787, 1:1000),donkey anti-goat IgG Alexa Fluor 488 (Invitrogen, A-11055, 1:500), donkey anti-goat IgG Alexa Fluor 555 (Invitrogen, A-32816, 1:500), donkey anti-goat IgG Alexa Fluor 647 (Invitrogen, A-21447, 1:500).

    Article Title: TRPV4 regulates β1 integrin-mediated cell-matrix adhesions and collagen remodeling.
    Article Snippet: GSK1016790A (a TRPV4 agonist), ionomycin (Ca2+ ionophore), bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma.bovine serum albumin (BSA), and PIPES were obtained from Millipore Sigma. ... The source of information for primary antibodies used is as follows: TRPV4 (ab39260 and ab191580, Abcam; MABS466, Millipore); β1- integrin (ab1952 and MAB1997, Millipore; clone KMI6, 14- 0292- 82, Invitrogen; clone 9EG7, 553 715, BD Biosciences); β3integrin (13166S, Cell Signaling); β5- integrin (3629S, Cell Signaling); α2- integrin (ab181548, Abcam); α11- integrin (ab198826, Abcam); α5- integrin (sc10729, Santa Cruz Biotechnology); αV- integrin (BD611012, BD Biosciences); β- actin (A1978, Millipore); DDR1 (sc- 374618, Santa Cruz Biotechnology); DDR2 (AF2538, R&D systems); Na+/K+ATPase α1 (3010S, Cell Signaling); GAPDH (MAB374, Millipore); Paxillin (ab32084, Abcam); Vimentin (NB300- 223, Novus Biologicals); Collagen I (PA5- 95137, Invitrogen); GFP (A- 11122, Invitrogen).. IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from LI- COR: Goat anti- Mouse (926- 68070; 926- 32210); Goat anti- Rabbit (926- 68071; 926- 32211); Donkey anti- Goat (926- 32214).IRDye® 680RD and 800CW secondary antibodies for immunoblotting were obtained from..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).. Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to the protocol provided. siRNA transfection Cells with > 80% sub-confluency were transfected with β-Catenin siRNA using Lipofectamine RNAiMAX (Invitrogen).Mouse serum ELISA experiment The levels of ANP and BNP in mouse serum were detected by Mouse ANP ELISA kit (Abcam, ab267800) and Mouse BNP ELISA kit (Novus, NBP2-70,011) according to..

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure.
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.As for quantification of immunostaining images, we used Image J to analyze the images from 3–5 different sections.. Primary antibodies were Vimentin (1:200, abcam, ab8978), DDR2 (1:200, R&D, MAB25381), PDGFRA (1:100, R&D, AF1062), POSTN (1:100, R&D, AF2955), THBS4 (1:100, R&D, AF2390), CD34 (1:100, abcam, ab81289), CD31 (1:100, R&D, AF3628), RFP antibody (1:300, Rockland, 600–401-379).. Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).Alexa Fluor-conjugated secondary antibodies used in this study included Donkey anti-Mouse IgG (1:500, Invitrogen, A21202 for Alexa Fluor 488), Donkey anti-Goat IgG (1:500, Invitrogen, A11055 for Alexa Fluor 488, A21447 for Alexa Fluor 647), Donkey anti-Rabbit IgG (1:500, Invitrogen, A31572 for Alexa Fluor 555).

    Article Title: Single cell and lineage tracing studies reveal the impact of CD34 + cells on myocardial fibrosis during heart failure
    Article Snippet: Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).Cells were lysed in RIPA lysis buffer supplemented with phosphatase inhibitor tablets (Roche, 04,906,837,001) and protease inhibitor tablet (Roche, 05,892,970,001).. Primary antibodies were Vimentin (Cell Signaling, 5741,1:1000), DDR2 (R&D, MAB25381, 1:2000), PDGFRA (R&D, AF1062, 1:1000), Periostin (R&D, AF2955, 1:1000), Wnt5a (Cell Signaling, 2530, 1:1000), β-Actin (Cell Signaling, 4970, 1:2000), β-Catenin (Cell Signaling, 8480, 1:2000), α-SMA (Sigma, A5228, 1:2000), Fibronectin (Cell Signaling, 26,836, 1:1000), Collagen I (abcam, ab6308, 1:1000), GAPDH (Cell Signaling, 5174, 1:2000), Smad2 (Cell Signaling, 5339, 1:1000), Phospho- Smad2 (Cell Signaling, 3108, 1:1000).



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    MAPK inhibitor-resistant melanoma cells exhibit enhanced DDR1/2 signaling. ( A ) Clonogenic assay showing the validation of drug-resistant cell lines. ( B ) Quantification of clonogenic assay shown in Fig. 1A. Data are presented as relative colony formation (%) normalized to the control group and expressed as the mean ± SEM, n = 3 replicates. Statistical significance was determined by two-way ANOVA. * p < 0.05, # p < 0.001, † p < 0.001, ‡ p < 0.0001. ( C ) Cell viability assay of RAF and MEK inhibitors in parental cells and drug-resistant cells for 48 h. Data are presented as the mean ± SEM, n = 3 replicates. ( D ) Western blot analysis of pERK and pAKT signaling responses in parental and resistant melanoma cells following 48-h treatment with vehicle or the corresponding resistance-selecting drug(s) at 1 µM. ( E ) Bubble plot representing Gene Set Enrichment Analysis (GSEA) using hallmark gene set comparing parental and resistant melanoma cells. Pathway enrichment was calculated using GSEA, with color indicating NES (blue = downregulated, red = upregulated) and bubble size representing −log10(p-value). ( F ) SK-MEL-2 and SK-MEL-2BR cells were stimulated with type I collagen (40 µg/ml) for 18 h to activate DDR signaling

    Journal: Cancer Cell International

    Article Title: PHI-501, a dual inhibitor of RAF and DDR1/2, overcomes MAPK drug resistance in Melanoma

    doi: 10.1186/s12935-026-04271-w

    Figure Lengend Snippet: MAPK inhibitor-resistant melanoma cells exhibit enhanced DDR1/2 signaling. ( A ) Clonogenic assay showing the validation of drug-resistant cell lines. ( B ) Quantification of clonogenic assay shown in Fig. 1A. Data are presented as relative colony formation (%) normalized to the control group and expressed as the mean ± SEM, n = 3 replicates. Statistical significance was determined by two-way ANOVA. * p < 0.05, # p < 0.001, † p < 0.001, ‡ p < 0.0001. ( C ) Cell viability assay of RAF and MEK inhibitors in parental cells and drug-resistant cells for 48 h. Data are presented as the mean ± SEM, n = 3 replicates. ( D ) Western blot analysis of pERK and pAKT signaling responses in parental and resistant melanoma cells following 48-h treatment with vehicle or the corresponding resistance-selecting drug(s) at 1 µM. ( E ) Bubble plot representing Gene Set Enrichment Analysis (GSEA) using hallmark gene set comparing parental and resistant melanoma cells. Pathway enrichment was calculated using GSEA, with color indicating NES (blue = downregulated, red = upregulated) and bubble size representing −log10(p-value). ( F ) SK-MEL-2 and SK-MEL-2BR cells were stimulated with type I collagen (40 µg/ml) for 18 h to activate DDR signaling

    Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies diluted in 5% BSA in TBST as indicated in the text: p-DDR1 (Cell Signaling Technology, CST14531 ), p-DDR1/2 (R&D Systems, MAB25382), p-AKT (Cell Signaling Technology, CST4058), AKT (Cell Signaling Technology, CST9272), p-MEK (Cell Signaling Technology, CST9121), p-ERK (Cell Signaling Technology, CST9101), ERK (Cell Signaling Technology, CST9102), Cyclin D1(Cell Signaling Technology, CST2978), and Survivin (Cell Signaling Technology, CST2808).

    Techniques: Clonogenic Assay, Biomarker Discovery, Control, Viability Assay, Western Blot

    DDR1 and DDR2 are associated with MAPK pathway activation and therapeutic resistance in melanoma. ( A ) Kaplan–Meier survival analysis showing patient outcomes based on DDR1 and DDR2 expression levels across all TCGA cancer types. Patients were stratified into high (top 25%) and low (bottom 25%) expression groups, and overall survival (OS) was analyzed. ( B ) Proteomic analysis of BRAF/NRAS-mutant SKCM. Box plots demonstrate differential expression of AKT and phosphorylated MEK1 (pMEK1-S217/S221) between wild-type and mutant groups. ( C ) Correlation heatmaps between DDR1/2 and key genes in the MAPK and AKT signaling pathways in TCGA-SKCM samples. Color intensity represents the correlation value, with red indicating a positive correlation. Significance levels are indicated by asterisks (* p < 0.05, ** p < 0.01, *** p < 0.001). ( D ) Correlation analysis revealing DDR1 expression (DepMap) and MAPK pathway inhibitor sensitivity (GDSC2) in melanoma cell lines. Scatter plots depicting correlations between DDR1 expression levels and IC50 to Dabrafenib (left) and Trametinib (right)

    Journal: Cancer Cell International

    Article Title: PHI-501, a dual inhibitor of RAF and DDR1/2, overcomes MAPK drug resistance in Melanoma

    doi: 10.1186/s12935-026-04271-w

    Figure Lengend Snippet: DDR1 and DDR2 are associated with MAPK pathway activation and therapeutic resistance in melanoma. ( A ) Kaplan–Meier survival analysis showing patient outcomes based on DDR1 and DDR2 expression levels across all TCGA cancer types. Patients were stratified into high (top 25%) and low (bottom 25%) expression groups, and overall survival (OS) was analyzed. ( B ) Proteomic analysis of BRAF/NRAS-mutant SKCM. Box plots demonstrate differential expression of AKT and phosphorylated MEK1 (pMEK1-S217/S221) between wild-type and mutant groups. ( C ) Correlation heatmaps between DDR1/2 and key genes in the MAPK and AKT signaling pathways in TCGA-SKCM samples. Color intensity represents the correlation value, with red indicating a positive correlation. Significance levels are indicated by asterisks (* p < 0.05, ** p < 0.01, *** p < 0.001). ( D ) Correlation analysis revealing DDR1 expression (DepMap) and MAPK pathway inhibitor sensitivity (GDSC2) in melanoma cell lines. Scatter plots depicting correlations between DDR1 expression levels and IC50 to Dabrafenib (left) and Trametinib (right)

    Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies diluted in 5% BSA in TBST as indicated in the text: p-DDR1 (Cell Signaling Technology, CST14531 ), p-DDR1/2 (R&D Systems, MAB25382), p-AKT (Cell Signaling Technology, CST4058), AKT (Cell Signaling Technology, CST9272), p-MEK (Cell Signaling Technology, CST9121), p-ERK (Cell Signaling Technology, CST9101), ERK (Cell Signaling Technology, CST9102), Cyclin D1(Cell Signaling Technology, CST2978), and Survivin (Cell Signaling Technology, CST2808).

    Techniques: Activation Assay, Expressing, Mutagenesis, Quantitative Proteomics, Protein-Protein interactions

    (A) CBF assessed by LSCI in WT, APP/PS1, and astrocyte specific Ddr2 overexpressing APP/PS1 mice. Left: Representative pseudocolor perfusion maps. Right: Quantification of relative perfusion units (Mean ± SEM; ** p < 0.01, one-way ANOVA). (B) Correlation between CBF (from A) and cortical Ddr2 mRNA levels (qPCR) across individual mice (Pearson r = −0.6081, p < 0.01). (C) In vivo (top) and ex vivo (bottom) whole brain fluorescence imaging after intravenous injection of the DDR2 targeting probe 1A12-mCherry. (D) Validation of 1A12-mCherry brain delivery and target specificity: Ex vivo brain sections for intrinsic mCherry fluorescence (red), anti-His tag immunofluorescence staining (green), and anti-DDR2 antibody HL2 staining (purple), the areas outlined by white squares are magnified in the adjacent panels. Scale bar: 30 μm for original images and 10 μm for enlarged images. (E) Schematic of the sequential probe injection protocol for vascular perfusion assessment: 1A12-mCherry followed 40 min later by Dextran-FITC (70 kDa). (F) Whole brain fluorescence imaging of vascular perfusion with Dextran-FITC (70 kDa). (G) Two photon microscopy of cortical vasculature. Representative images show mCherry signal (red) and dextran-FITC vasculature (green) in WT, APP/PS1, and APP/PS1-DDR2 mice, scale bars: 50 μm. (H) Representative two-photon microscopy images of vascular leakage after injection of dextran-FITC (4 kDa, green) and dextran-RB (70 kDa, red), scale bars: 50 μm. (I) Ventricular morphology analyzed by MRI based volumetric reconstruction. Left: Representative images of lateral ventricles from each group. Right: Quantification of lateral ventricular volume (Mean ± SEM; * p < 0.05, ** p < 0.01, one-way ANOVA). (J) CSF flow assessed by cisterna magna injection of high molecular weight dextran (70 kDa, green). Fluorescence stereo microscope images displaying the spatial distribution of the glymphatic system (green) in the brains ( top row ), and corresponding whole brain fluorescence imaging system scans of the same brains( bottom row ), scale bars: 2 mm.

    Journal: medRxiv

    Article Title: A brain-persistent DDR2-degrading antibody reverses Alzheimer’s pathologies by restoring brain fluid dynamics and metabolic clearance

    doi: 10.64898/2026.03.17.26348575

    Figure Lengend Snippet: (A) CBF assessed by LSCI in WT, APP/PS1, and astrocyte specific Ddr2 overexpressing APP/PS1 mice. Left: Representative pseudocolor perfusion maps. Right: Quantification of relative perfusion units (Mean ± SEM; ** p < 0.01, one-way ANOVA). (B) Correlation between CBF (from A) and cortical Ddr2 mRNA levels (qPCR) across individual mice (Pearson r = −0.6081, p < 0.01). (C) In vivo (top) and ex vivo (bottom) whole brain fluorescence imaging after intravenous injection of the DDR2 targeting probe 1A12-mCherry. (D) Validation of 1A12-mCherry brain delivery and target specificity: Ex vivo brain sections for intrinsic mCherry fluorescence (red), anti-His tag immunofluorescence staining (green), and anti-DDR2 antibody HL2 staining (purple), the areas outlined by white squares are magnified in the adjacent panels. Scale bar: 30 μm for original images and 10 μm for enlarged images. (E) Schematic of the sequential probe injection protocol for vascular perfusion assessment: 1A12-mCherry followed 40 min later by Dextran-FITC (70 kDa). (F) Whole brain fluorescence imaging of vascular perfusion with Dextran-FITC (70 kDa). (G) Two photon microscopy of cortical vasculature. Representative images show mCherry signal (red) and dextran-FITC vasculature (green) in WT, APP/PS1, and APP/PS1-DDR2 mice, scale bars: 50 μm. (H) Representative two-photon microscopy images of vascular leakage after injection of dextran-FITC (4 kDa, green) and dextran-RB (70 kDa, red), scale bars: 50 μm. (I) Ventricular morphology analyzed by MRI based volumetric reconstruction. Left: Representative images of lateral ventricles from each group. Right: Quantification of lateral ventricular volume (Mean ± SEM; * p < 0.05, ** p < 0.01, one-way ANOVA). (J) CSF flow assessed by cisterna magna injection of high molecular weight dextran (70 kDa, green). Fluorescence stereo microscope images displaying the spatial distribution of the glymphatic system (green) in the brains ( top row ), and corresponding whole brain fluorescence imaging system scans of the same brains( bottom row ), scale bars: 2 mm.

    Article Snippet: The primary antibodies used in this study were as follows: anti-DDR2 (1:1000; Servicebio, GB112568), anti-DDR2 (1:1000, R&D, AF2538), anti-BACE1 (1:50; Cell Signaling Technology, no. 5606), anti-Collagen IV (1:1000; abcam, ab6586), anti-PDGFRβ (1:1000; Cell Signaling Technology, no. 3169), anti-Occludin (1:1000; Cell Signaling Technology, no. 91131), anti-ZO1 (1:1000; abcam, ab276131), anti-β-actin (1:1000; Yeasen, 30102ES60).

    Techniques: In Vivo, Ex Vivo, Fluorescence, Imaging, Injection, Biomarker Discovery, Immunofluorescence, Staining, Microscopy, High Molecular Weight