Review



imagequant data analysis software  (Danaher Inc)


Bioz Verified Symbol Danaher Inc is a verified supplier
Bioz Manufacturer Symbol Danaher Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Danaher Inc imagequant data analysis software
    Imagequant Data Analysis Software, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/imagequant+analysis+software/us12516375-1245-66-70
    Average 86 stars, based on 1 article reviews
    imagequant data analysis software - by Bioz Stars, 2026-10
    86/100 stars

    Images

    Related Articles

    Incubation:

    Article Title: Compositions, devices, systems, and methods for using a nanopore
    Article Snippet: .. After incubation at 65° C. for 18 hours, the hybridization buffer is removed, and the substrate is washed sequentially under increasingly stringent conditions, up to 40 mM sodium phosphate, 1% SDS, 1 mM EDTA at 65° C. To detect signal produced by a radiolabeled probe hybridized on a membrane, the substrate is exposed to a PHOSPHORIMAGER cassette (Amersham Pharmacia Biotech), and the image is analyzed using IMAGEQUANT data analysis software (Amersham Pharmacia Biotech). ..

    Hybridization:

    Article Title: Compositions, devices, systems, and methods for using a nanopore
    Article Snippet: .. After incubation at 65° C. for 18 hours, the hybridization buffer is removed, and the substrate is washed sequentially under increasingly stringent conditions, up to 40 mM sodium phosphate, 1% SDS, 1 mM EDTA at 65° C. To detect signal produced by a radiolabeled probe hybridized on a membrane, the substrate is exposed to a PHOSPHORIMAGER cassette (Amersham Pharmacia Biotech), and the image is analyzed using IMAGEQUANT data analysis software (Amersham Pharmacia Biotech). ..

    Produced:

    Article Title: Compositions, devices, systems, and methods for using a nanopore
    Article Snippet: .. After incubation at 65° C. for 18 hours, the hybridization buffer is removed, and the substrate is washed sequentially under increasingly stringent conditions, up to 40 mM sodium phosphate, 1% SDS, 1 mM EDTA at 65° C. To detect signal produced by a radiolabeled probe hybridized on a membrane, the substrate is exposed to a PHOSPHORIMAGER cassette (Amersham Pharmacia Biotech), and the image is analyzed using IMAGEQUANT data analysis software (Amersham Pharmacia Biotech). ..

    Membrane:

    Article Title: Compositions, devices, systems, and methods for using a nanopore
    Article Snippet: .. After incubation at 65° C. for 18 hours, the hybridization buffer is removed, and the substrate is washed sequentially under increasingly stringent conditions, up to 40 mM sodium phosphate, 1% SDS, 1 mM EDTA at 65° C. To detect signal produced by a radiolabeled probe hybridized on a membrane, the substrate is exposed to a PHOSPHORIMAGER cassette (Amersham Pharmacia Biotech), and the image is analyzed using IMAGEQUANT data analysis software (Amersham Pharmacia Biotech). ..

    Software:

    Article Title: Compositions, devices, systems, and methods for using a nanopore
    Article Snippet: .. After incubation at 65° C. for 18 hours, the hybridization buffer is removed, and the substrate is washed sequentially under increasingly stringent conditions, up to 40 mM sodium phosphate, 1% SDS, 1 mM EDTA at 65° C. To detect signal produced by a radiolabeled probe hybridized on a membrane, the substrate is exposed to a PHOSPHORIMAGER cassette (Amersham Pharmacia Biotech), and the image is analyzed using IMAGEQUANT data analysis software (Amersham Pharmacia Biotech). ..

    Article Title: Potential of antiviral drug oseltamivir for the treatment of liver cancer
    Article Snippet: Next, the secondary antibodies conjugated with horse-radish peroxidase (HRP) (1:5,000; cat. no. sc-2004 or sc-2005; Santa Cruz Biotechnology, Inc.) were added and incubated for 1 h at 25°C. .. To detect the antigen-antibody complexes, Immobilion Western HRP Chemiluminescent Substrate (EMD Millipore) and a densitometry apparatus LAS-4000 (Image Analysis Software: GE ImageQuant TL 8.1; GE Healthcare Life Sciences) were used. .. In addition, an autophagy inhibitor, chloroquine diphosphate (CQ; cat. no. L10382; LC3B Antibody Kit; Invitrogen; Thermo Fisher Scientific Inc.), was used to verify the participation of the autophagic mechanism to oseltamivir-induced death in Huh-7 and HepG2 cells.

    Article Title: Vitro evolution in microfluidic systems
    Article Snippet: .. DNA is visualized by staining with ethidium bromide and quantified using ImageQuant TL gel analysis software (Amersham Biosciences). ..

    Article Title: Design, synthesis and biological evaluation of novel orthosteric-allosteric ligands of the cannabinoid receptor type 2 (CB 2 R)
    Article Snippet: Next, the membranes were incubated with HRP-labeled secondary anti-rabbit (#MAB201P, Merck-Millipore, Darmstadt, Germany) for 2 h at room temperature. .. Detection of chemiluminescence signals and densitometric analysis of blots were performed using ImageQuant LAS 4000 (GE Healthcare, Milan, Italy) and ImageLab software (Bio-Rad, Hercules, CA, United States ), respectively. ..

    Article Title: Flexible linker modulates the binding affinity of the TP901-1 CI phage repressor to DNA.
    Article Snippet: doi:10.1111/febs.16238 Temperate bacteriophages can switch between two life cycles following infection of a host bacterium: the lytic or lysogenic life cycle.. The choice between these is controlled by a bistable genetic switch.. We investigated the genetic switch of the lactococcal temperate bacteriophage, TP901-1, which is controlled by two regulatory proteins, the Clear 1 (CI) repressor and modulator of repression (MOR) antirepressor.

    Article Title: Mevalonate Kinase Deficiency and Squalene Synthase Inhibitor (TAK-475): The Balance to Extinguish the Inflammation
    Article Snippet: After incubation with secondary antibodies (anti-mouse or -rabbit IgG HRP-conjugated; Sigma-Aldrich, St. Louis, MO, USA), a specific band detection was performed with the WesternBright Quantum kit (Advansta, Menlo Park, CA, USA). .. Images’ acquisition and analysis were performed using the ImageQuantTM LAS 4000 imager and TL software (GE Healthcare, Buckinghamshire, UK). ..

    Article Title: Molecular definition of the interaction between a tumor-specific tetrabranched peptide and LRP6 receptor.
    Article Snippet: The tumor-specific tetrabranched peptide NT4 binds membrane sulfate glycosaminoglycans and receptors belonging to the low density lipoprotein receptor-related protein (LRP) family, like LRP6, which are overexpressed in cancer.. The binding occurs through a multimeric positively-charged motif of NT4 that interacts with negatively charged motives in both glycosaminoglycans and LRP receptors.. LRP6 has an essential function in canonical Wnt signaling, acting together with receptors of the Frizzled family as coreceptor for Wnt ligands.

    Article Title: Rosmarinic Acid Inhibits Ultraviolet B-Mediated Oxidative Damage via the AKT/ERK-NRF2-GSH Pathway In Vitro and In Vivo .
    Article Snippet: .. The final mixture was resolved by electrophoresis on a 1% agarose gel, stained with ethidium bromide, and photographed under ultraviolet light using ImageQuant TL analysis software (Amersham Biosciences, Uppsala, Sweden). ..

    Western Blot:

    Article Title: Potential of antiviral drug oseltamivir for the treatment of liver cancer
    Article Snippet: Next, the secondary antibodies conjugated with horse-radish peroxidase (HRP) (1:5,000; cat. no. sc-2004 or sc-2005; Santa Cruz Biotechnology, Inc.) were added and incubated for 1 h at 25°C. .. To detect the antigen-antibody complexes, Immobilion Western HRP Chemiluminescent Substrate (EMD Millipore) and a densitometry apparatus LAS-4000 (Image Analysis Software: GE ImageQuant TL 8.1; GE Healthcare Life Sciences) were used. .. In addition, an autophagy inhibitor, chloroquine diphosphate (CQ; cat. no. L10382; LC3B Antibody Kit; Invitrogen; Thermo Fisher Scientific Inc.), was used to verify the participation of the autophagic mechanism to oseltamivir-induced death in Huh-7 and HepG2 cells.

    Staining:

    Article Title: Vitro evolution in microfluidic systems
    Article Snippet: .. DNA is visualized by staining with ethidium bromide and quantified using ImageQuant TL gel analysis software (Amersham Biosciences). ..

    Article Title: Rosmarinic Acid Inhibits Ultraviolet B-Mediated Oxidative Damage via the AKT/ERK-NRF2-GSH Pathway In Vitro and In Vivo .
    Article Snippet: .. The final mixture was resolved by electrophoresis on a 1% agarose gel, stained with ethidium bromide, and photographed under ultraviolet light using ImageQuant TL analysis software (Amersham Biosciences, Uppsala, Sweden). ..

    Analysis:

    Article Title: Molecular definition of the interaction between a tumor-specific tetrabranched peptide and LRP6 receptor.
    Article Snippet: The tumor-specific tetrabranched peptide NT4 binds membrane sulfate glycosaminoglycans and receptors belonging to the low density lipoprotein receptor-related protein (LRP) family, like LRP6, which are overexpressed in cancer.. The binding occurs through a multimeric positively-charged motif of NT4 that interacts with negatively charged motives in both glycosaminoglycans and LRP receptors.. LRP6 has an essential function in canonical Wnt signaling, acting together with receptors of the Frizzled family as coreceptor for Wnt ligands.

    Electrophoresis:

    Article Title: Rosmarinic Acid Inhibits Ultraviolet B-Mediated Oxidative Damage via the AKT/ERK-NRF2-GSH Pathway In Vitro and In Vivo .
    Article Snippet: .. The final mixture was resolved by electrophoresis on a 1% agarose gel, stained with ethidium bromide, and photographed under ultraviolet light using ImageQuant TL analysis software (Amersham Biosciences, Uppsala, Sweden). ..

    Agarose Gel Electrophoresis:

    Article Title: Rosmarinic Acid Inhibits Ultraviolet B-Mediated Oxidative Damage via the AKT/ERK-NRF2-GSH Pathway In Vitro and In Vivo .
    Article Snippet: .. The final mixture was resolved by electrophoresis on a 1% agarose gel, stained with ethidium bromide, and photographed under ultraviolet light using ImageQuant TL analysis software (Amersham Biosciences, Uppsala, Sweden). ..



    Similar Products

    86
    Cytiva Europe imagequant image analysis software
    Imagequant Image Analysis Software, supplied by Cytiva Europe, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/imagequant+analysis+software/pmc11344278-232-21-25
    Average 86 stars, based on 1 article reviews
    imagequant image analysis software - by Bioz Stars, 2026-10
    86/100 stars
      Buy from Supplier

    86
    Danaher Inc imagequant data analysis software
    Imagequant Data Analysis Software, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/imagequant+analysis+software/us12516375-1245-66-70
    Average 86 stars, based on 1 article reviews
    imagequant data analysis software - by Bioz Stars, 2026-10
    86/100 stars
      Buy from Supplier

    90
    Amersham Life Sciences Inc imagequant tl analysis software
    Imagequant Tl Analysis Software, supplied by Amersham Life Sciences Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/imagequant+analysis+software/imagequant+software/pm40249806-196-16-20
    Average 90 stars, based on 1 article reviews
    imagequant tl analysis software - by Bioz Stars, 2026-10
    90/100 stars
      Buy from Supplier

    90
    Amersham Life Sciences Inc imagequant™ tl 10.2 analysis software
    Imagequant™ Tl 10.2 Analysis Software, supplied by Amersham Life Sciences Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/imagequant+analysis+software/imagequant+tl+software/pmc11549352-71-0-6
    Average 90 stars, based on 1 article reviews
    imagequant™ tl 10.2 analysis software - by Bioz Stars, 2026-10
    90/100 stars
      Buy from Supplier

    90
    Cytiva Europe imagequant tl analysis software
    Phosphorylation of MSK1 via the p38 kinase pathway upon BMP-6 stimulation in ATDC5 cells. A , phosphorylation of MSK1 (Ser376) and MSK1 (Ser360) by BMP-6. ATDC5 cells were stimulated with 25 ng/ml BMP-6 for the indicated times. The total cell lysates were then used for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-ERK1/2 (Thr202/Tyr204), phospho-p38 (Thr180/Tyr182), phospho-JNK (Thr183/Tyr185), phospho-Smad1/5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6. B , Inhibition of BMP-6-induced MSK1 phosphorylation by dorsomorphin, a BMP type I receptor kinase inhibitor. ATDC5 cells were pretreated with 10 μM dorsomorphin for 1 h. Thereafter, cells were stimulated with 25 ng/ml BMP-6 for the indicated times. The total cell lysates were used for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of dorsomorphin. C , Involvement of the ERK pathway in the phosphorylation of MSK1 by BMP-6. ATDC5 cells were pretreated with 20 μM U0126 1 h before stimulation with 25 ng/ml BMP-6 for the indicated times. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-ERK1/2 (Thr202/Tyr204), ERK1/2, phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh, eighth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of U0126. D , decrease in BMP-6-induced phosphorylation of MSK1 in the presence of the p38 kinase inhibitor. ATDC5 cells pretreated with 10 μM SB203580, a p38 kinase inhibitor, for 1 h were stimulated with 25 ng/ml BMP-6 for the indicated times. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-p38 (Thr180/Tyr182), p38, phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh, eighth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of SB203580. A-D , white and black arrowheads indicate phospho-MSK1 (Ser376)-specific and nonspecific bands, respectively. E , decrease in BMP-6-mediated phospho-CREB (Ser133) expression by the MSK1 kinase inhibitor, SB747651A. ATDC5 cells were cultured with 10 μM SB747651A for 1 h. Prior to preparation of the total lysates, 25 ng/ml BMP-6 was added to the culture media for the indicated times. The total expression levels of phospho-CREB (Ser133), phospho-p38 (Thr180/Tyr182), phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth , and bottom panels , respectively. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of SB747651A. F , chondrogenic differentiation of ATDC5 cells by BMP-6 in the presence of kinase inhibitors. ATDC5 cells cultured at a high density were stimulated with 25 ng/ml BMP-6 in the presence (+) or absence (−) of kinase inhibitors (20 μM U0126, 10 μM SB203580, 10 μM SB747651A, or 10 μM dorsomorphin). Two weeks later, the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using <t>ImageQuant</t> TL. The relative intensity of the stained area was calculated with respect to the 1-h treatment of cells with BMP-6 alone. G , representative images of ex vivo chondrogenic differentiation of mouse metatarsal bones induced by BMP-6 in the presence of kinase inhibitors. Each mouse metatarsal bone was cultured with or without 25 ng/ml BMP-6 for 7 days, either in the absence or presence of kinase inhibitors (20 μM U0126, 10 μM SB203580, 10 μM SB747651A, or 10 μM dorsomorphin). Then, each bone was stained with Alcian blue and Alizarin Red S. Two representative images are shown in each treatment. a , proliferative chondrocytes; b , clear zone represented as hypertrophic chondrocytes; c , chondrocyte matrix calcified by mature hypertrophic chondrocytes stained with alizarin red S. H , relative area of chondrocyte differentiation in mouse metatarsal bones induced by BMP-6 in the presence of kinase inhibitors. The total areas of chondrocyte differentiation (a + b + c) were measured. The total areas of chondrocyte differentiation (a + b + c) were measured using ImageJ. Asterisks indicate significant differences. BMP, bone morphogenetic protein; CREB, cAMP-response element-binding protein; MSK, mitogen- and stress-activated protein kinase; ND; not determined.
    Imagequant Tl Analysis Software, supplied by Cytiva Europe, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/imagequant+analysis+software/imagequant+software/pmc11541777-329-8-12
    Average 90 stars, based on 1 article reviews
    imagequant tl analysis software - by Bioz Stars, 2026-10
    90/100 stars
      Buy from Supplier

    86
    Cytiva Europe imagequant analysis software
    Phosphorylation of MSK1 via the p38 kinase pathway upon BMP-6 stimulation in ATDC5 cells. A , phosphorylation of MSK1 (Ser376) and MSK1 (Ser360) by BMP-6. ATDC5 cells were stimulated with 25 ng/ml BMP-6 for the indicated times. The total cell lysates were then used for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-ERK1/2 (Thr202/Tyr204), phospho-p38 (Thr180/Tyr182), phospho-JNK (Thr183/Tyr185), phospho-Smad1/5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6. B , Inhibition of BMP-6-induced MSK1 phosphorylation by dorsomorphin, a BMP type I receptor kinase inhibitor. ATDC5 cells were pretreated with 10 μM dorsomorphin for 1 h. Thereafter, cells were stimulated with 25 ng/ml BMP-6 for the indicated times. The total cell lysates were used for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of dorsomorphin. C , Involvement of the ERK pathway in the phosphorylation of MSK1 by BMP-6. ATDC5 cells were pretreated with 20 μM U0126 1 h before stimulation with 25 ng/ml BMP-6 for the indicated times. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-ERK1/2 (Thr202/Tyr204), ERK1/2, phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh, eighth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of U0126. D , decrease in BMP-6-induced phosphorylation of MSK1 in the presence of the p38 kinase inhibitor. ATDC5 cells pretreated with 10 μM SB203580, a p38 kinase inhibitor, for 1 h were stimulated with 25 ng/ml BMP-6 for the indicated times. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-p38 (Thr180/Tyr182), p38, phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh, eighth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of SB203580. A-D , white and black arrowheads indicate phospho-MSK1 (Ser376)-specific and nonspecific bands, respectively. E , decrease in BMP-6-mediated phospho-CREB (Ser133) expression by the MSK1 kinase inhibitor, SB747651A. ATDC5 cells were cultured with 10 μM SB747651A for 1 h. Prior to preparation of the total lysates, 25 ng/ml BMP-6 was added to the culture media for the indicated times. The total expression levels of phospho-CREB (Ser133), phospho-p38 (Thr180/Tyr182), phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth , and bottom panels , respectively. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of SB747651A. F , chondrogenic differentiation of ATDC5 cells by BMP-6 in the presence of kinase inhibitors. ATDC5 cells cultured at a high density were stimulated with 25 ng/ml BMP-6 in the presence (+) or absence (−) of kinase inhibitors (20 μM U0126, 10 μM SB203580, 10 μM SB747651A, or 10 μM dorsomorphin). Two weeks later, the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using <t>ImageQuant</t> TL. The relative intensity of the stained area was calculated with respect to the 1-h treatment of cells with BMP-6 alone. G , representative images of ex vivo chondrogenic differentiation of mouse metatarsal bones induced by BMP-6 in the presence of kinase inhibitors. Each mouse metatarsal bone was cultured with or without 25 ng/ml BMP-6 for 7 days, either in the absence or presence of kinase inhibitors (20 μM U0126, 10 μM SB203580, 10 μM SB747651A, or 10 μM dorsomorphin). Then, each bone was stained with Alcian blue and Alizarin Red S. Two representative images are shown in each treatment. a , proliferative chondrocytes; b , clear zone represented as hypertrophic chondrocytes; c , chondrocyte matrix calcified by mature hypertrophic chondrocytes stained with alizarin red S. H , relative area of chondrocyte differentiation in mouse metatarsal bones induced by BMP-6 in the presence of kinase inhibitors. The total areas of chondrocyte differentiation (a + b + c) were measured. The total areas of chondrocyte differentiation (a + b + c) were measured using ImageJ. Asterisks indicate significant differences. BMP, bone morphogenetic protein; CREB, cAMP-response element-binding protein; MSK, mitogen- and stress-activated protein kinase; ND; not determined.
    Imagequant Analysis Software, supplied by Cytiva Europe, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/imagequant+analysis+software/10__1113_slash_jp286861-120-22-25
    Average 86 stars, based on 1 article reviews
    imagequant analysis software - by Bioz Stars, 2026-10
    86/100 stars
      Buy from Supplier

    86
    Cytiva Europe imagequant tl image analysis software
    Phosphorylation of MSK1 via the p38 kinase pathway upon BMP-6 stimulation in ATDC5 cells. A , phosphorylation of MSK1 (Ser376) and MSK1 (Ser360) by BMP-6. ATDC5 cells were stimulated with 25 ng/ml BMP-6 for the indicated times. The total cell lysates were then used for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-ERK1/2 (Thr202/Tyr204), phospho-p38 (Thr180/Tyr182), phospho-JNK (Thr183/Tyr185), phospho-Smad1/5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6. B , Inhibition of BMP-6-induced MSK1 phosphorylation by dorsomorphin, a BMP type I receptor kinase inhibitor. ATDC5 cells were pretreated with 10 μM dorsomorphin for 1 h. Thereafter, cells were stimulated with 25 ng/ml BMP-6 for the indicated times. The total cell lysates were used for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of dorsomorphin. C , Involvement of the ERK pathway in the phosphorylation of MSK1 by BMP-6. ATDC5 cells were pretreated with 20 μM U0126 1 h before stimulation with 25 ng/ml BMP-6 for the indicated times. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-ERK1/2 (Thr202/Tyr204), ERK1/2, phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh, eighth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of U0126. D , decrease in BMP-6-induced phosphorylation of MSK1 in the presence of the p38 kinase inhibitor. ATDC5 cells pretreated with 10 μM SB203580, a p38 kinase inhibitor, for 1 h were stimulated with 25 ng/ml BMP-6 for the indicated times. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-p38 (Thr180/Tyr182), p38, phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh, eighth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of SB203580. A-D , white and black arrowheads indicate phospho-MSK1 (Ser376)-specific and nonspecific bands, respectively. E , decrease in BMP-6-mediated phospho-CREB (Ser133) expression by the MSK1 kinase inhibitor, SB747651A. ATDC5 cells were cultured with 10 μM SB747651A for 1 h. Prior to preparation of the total lysates, 25 ng/ml BMP-6 was added to the culture media for the indicated times. The total expression levels of phospho-CREB (Ser133), phospho-p38 (Thr180/Tyr182), phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth , and bottom panels , respectively. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of SB747651A. F , chondrogenic differentiation of ATDC5 cells by BMP-6 in the presence of kinase inhibitors. ATDC5 cells cultured at a high density were stimulated with 25 ng/ml BMP-6 in the presence (+) or absence (−) of kinase inhibitors (20 μM U0126, 10 μM SB203580, 10 μM SB747651A, or 10 μM dorsomorphin). Two weeks later, the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using <t>ImageQuant</t> TL. The relative intensity of the stained area was calculated with respect to the 1-h treatment of cells with BMP-6 alone. G , representative images of ex vivo chondrogenic differentiation of mouse metatarsal bones induced by BMP-6 in the presence of kinase inhibitors. Each mouse metatarsal bone was cultured with or without 25 ng/ml BMP-6 for 7 days, either in the absence or presence of kinase inhibitors (20 μM U0126, 10 μM SB203580, 10 μM SB747651A, or 10 μM dorsomorphin). Then, each bone was stained with Alcian blue and Alizarin Red S. Two representative images are shown in each treatment. a , proliferative chondrocytes; b , clear zone represented as hypertrophic chondrocytes; c , chondrocyte matrix calcified by mature hypertrophic chondrocytes stained with alizarin red S. H , relative area of chondrocyte differentiation in mouse metatarsal bones induced by BMP-6 in the presence of kinase inhibitors. The total areas of chondrocyte differentiation (a + b + c) were measured. The total areas of chondrocyte differentiation (a + b + c) were measured using ImageJ. Asterisks indicate significant differences. BMP, bone morphogenetic protein; CREB, cAMP-response element-binding protein; MSK, mitogen- and stress-activated protein kinase; ND; not determined.
    Imagequant Tl Image Analysis Software, supplied by Cytiva Europe, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/imagequant+analysis+software/pmc10980023-533-12-17
    Average 86 stars, based on 1 article reviews
    imagequant tl image analysis software - by Bioz Stars, 2026-10
    86/100 stars
      Buy from Supplier

    Image Search Results


    Phosphorylation of MSK1 via the p38 kinase pathway upon BMP-6 stimulation in ATDC5 cells. A , phosphorylation of MSK1 (Ser376) and MSK1 (Ser360) by BMP-6. ATDC5 cells were stimulated with 25 ng/ml BMP-6 for the indicated times. The total cell lysates were then used for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-ERK1/2 (Thr202/Tyr204), phospho-p38 (Thr180/Tyr182), phospho-JNK (Thr183/Tyr185), phospho-Smad1/5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6. B , Inhibition of BMP-6-induced MSK1 phosphorylation by dorsomorphin, a BMP type I receptor kinase inhibitor. ATDC5 cells were pretreated with 10 μM dorsomorphin for 1 h. Thereafter, cells were stimulated with 25 ng/ml BMP-6 for the indicated times. The total cell lysates were used for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of dorsomorphin. C , Involvement of the ERK pathway in the phosphorylation of MSK1 by BMP-6. ATDC5 cells were pretreated with 20 μM U0126 1 h before stimulation with 25 ng/ml BMP-6 for the indicated times. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-ERK1/2 (Thr202/Tyr204), ERK1/2, phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh, eighth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of U0126. D , decrease in BMP-6-induced phosphorylation of MSK1 in the presence of the p38 kinase inhibitor. ATDC5 cells pretreated with 10 μM SB203580, a p38 kinase inhibitor, for 1 h were stimulated with 25 ng/ml BMP-6 for the indicated times. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-p38 (Thr180/Tyr182), p38, phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh, eighth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of SB203580. A-D , white and black arrowheads indicate phospho-MSK1 (Ser376)-specific and nonspecific bands, respectively. E , decrease in BMP-6-mediated phospho-CREB (Ser133) expression by the MSK1 kinase inhibitor, SB747651A. ATDC5 cells were cultured with 10 μM SB747651A for 1 h. Prior to preparation of the total lysates, 25 ng/ml BMP-6 was added to the culture media for the indicated times. The total expression levels of phospho-CREB (Ser133), phospho-p38 (Thr180/Tyr182), phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth , and bottom panels , respectively. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of SB747651A. F , chondrogenic differentiation of ATDC5 cells by BMP-6 in the presence of kinase inhibitors. ATDC5 cells cultured at a high density were stimulated with 25 ng/ml BMP-6 in the presence (+) or absence (−) of kinase inhibitors (20 μM U0126, 10 μM SB203580, 10 μM SB747651A, or 10 μM dorsomorphin). Two weeks later, the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using ImageQuant TL. The relative intensity of the stained area was calculated with respect to the 1-h treatment of cells with BMP-6 alone. G , representative images of ex vivo chondrogenic differentiation of mouse metatarsal bones induced by BMP-6 in the presence of kinase inhibitors. Each mouse metatarsal bone was cultured with or without 25 ng/ml BMP-6 for 7 days, either in the absence or presence of kinase inhibitors (20 μM U0126, 10 μM SB203580, 10 μM SB747651A, or 10 μM dorsomorphin). Then, each bone was stained with Alcian blue and Alizarin Red S. Two representative images are shown in each treatment. a , proliferative chondrocytes; b , clear zone represented as hypertrophic chondrocytes; c , chondrocyte matrix calcified by mature hypertrophic chondrocytes stained with alizarin red S. H , relative area of chondrocyte differentiation in mouse metatarsal bones induced by BMP-6 in the presence of kinase inhibitors. The total areas of chondrocyte differentiation (a + b + c) were measured. The total areas of chondrocyte differentiation (a + b + c) were measured using ImageJ. Asterisks indicate significant differences. BMP, bone morphogenetic protein; CREB, cAMP-response element-binding protein; MSK, mitogen- and stress-activated protein kinase; ND; not determined.

    Journal: The Journal of Biological Chemistry

    Article Title: Involvement of mitogen- and stress-activated protein kinase 1 in BMP-6–induced chondrocyte differentiation

    doi: 10.1016/j.jbc.2024.107806

    Figure Lengend Snippet: Phosphorylation of MSK1 via the p38 kinase pathway upon BMP-6 stimulation in ATDC5 cells. A , phosphorylation of MSK1 (Ser376) and MSK1 (Ser360) by BMP-6. ATDC5 cells were stimulated with 25 ng/ml BMP-6 for the indicated times. The total cell lysates were then used for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-ERK1/2 (Thr202/Tyr204), phospho-p38 (Thr180/Tyr182), phospho-JNK (Thr183/Tyr185), phospho-Smad1/5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6. B , Inhibition of BMP-6-induced MSK1 phosphorylation by dorsomorphin, a BMP type I receptor kinase inhibitor. ATDC5 cells were pretreated with 10 μM dorsomorphin for 1 h. Thereafter, cells were stimulated with 25 ng/ml BMP-6 for the indicated times. The total cell lysates were used for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of dorsomorphin. C , Involvement of the ERK pathway in the phosphorylation of MSK1 by BMP-6. ATDC5 cells were pretreated with 20 μM U0126 1 h before stimulation with 25 ng/ml BMP-6 for the indicated times. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-ERK1/2 (Thr202/Tyr204), ERK1/2, phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh, eighth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of U0126. D , decrease in BMP-6-induced phosphorylation of MSK1 in the presence of the p38 kinase inhibitor. ATDC5 cells pretreated with 10 μM SB203580, a p38 kinase inhibitor, for 1 h were stimulated with 25 ng/ml BMP-6 for the indicated times. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-p38 (Thr180/Tyr182), p38, phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh, eighth , and bottom panels , respectively. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of SB203580. A-D , white and black arrowheads indicate phospho-MSK1 (Ser376)-specific and nonspecific bands, respectively. E , decrease in BMP-6-mediated phospho-CREB (Ser133) expression by the MSK1 kinase inhibitor, SB747651A. ATDC5 cells were cultured with 10 μM SB747651A for 1 h. Prior to preparation of the total lysates, 25 ng/ml BMP-6 was added to the culture media for the indicated times. The total expression levels of phospho-CREB (Ser133), phospho-p38 (Thr180/Tyr182), phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth , and bottom panels , respectively. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells with BMP-6 in the absence of SB747651A. F , chondrogenic differentiation of ATDC5 cells by BMP-6 in the presence of kinase inhibitors. ATDC5 cells cultured at a high density were stimulated with 25 ng/ml BMP-6 in the presence (+) or absence (−) of kinase inhibitors (20 μM U0126, 10 μM SB203580, 10 μM SB747651A, or 10 μM dorsomorphin). Two weeks later, the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using ImageQuant TL. The relative intensity of the stained area was calculated with respect to the 1-h treatment of cells with BMP-6 alone. G , representative images of ex vivo chondrogenic differentiation of mouse metatarsal bones induced by BMP-6 in the presence of kinase inhibitors. Each mouse metatarsal bone was cultured with or without 25 ng/ml BMP-6 for 7 days, either in the absence or presence of kinase inhibitors (20 μM U0126, 10 μM SB203580, 10 μM SB747651A, or 10 μM dorsomorphin). Then, each bone was stained with Alcian blue and Alizarin Red S. Two representative images are shown in each treatment. a , proliferative chondrocytes; b , clear zone represented as hypertrophic chondrocytes; c , chondrocyte matrix calcified by mature hypertrophic chondrocytes stained with alizarin red S. H , relative area of chondrocyte differentiation in mouse metatarsal bones induced by BMP-6 in the presence of kinase inhibitors. The total areas of chondrocyte differentiation (a + b + c) were measured. The total areas of chondrocyte differentiation (a + b + c) were measured using ImageJ. Asterisks indicate significant differences. BMP, bone morphogenetic protein; CREB, cAMP-response element-binding protein; MSK, mitogen- and stress-activated protein kinase; ND; not determined.

    Article Snippet: The intensity of each band was measured using ImageQuant TL analysis software (Cytiva) after each gel image was captured with the ImageQuant LAS 4000 mini (Amersham).

    Techniques: Phospho-proteomics, Western Blot, Expressing, Inhibition, Cell Culture, Staining, Ex Vivo, Binding Assay

    I nflu ence of MSK1 deficiency on BMP-6–mediated chondrogenic differentiation. A , expression of MSK1 in ATDC5 cells. Total lysates from each transformant were prepared for Western blot analyses. The total expression levels of MSK1 and β-actin are indicated in the upper and lower panels , respectively. White and black arrowheads indicate MSK1-specific and nonspecific bands, respectively. The intensity of the band for MSK1 was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to cells carrying shControl. B , chondrogenic differentiation of ATDC5 cells with decreased expression of MSK1 in the presence of BMP-6. The cells were seeded at a high density and then stimulated with 25 ng/ml BMP-6. Seven ( upper panels ) or 14 days later ( lower panels ), the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using ImageQuant TL. The relative intensity of the stained area was calculated with respect to the cells carrying shControl with BMP-6 for 14 days. BMP, bone morphogenetic protein; MSK, mitogen- and stress-activated protein kinase; qPCR, quantitative PCR.

    Journal: The Journal of Biological Chemistry

    Article Title: Involvement of mitogen- and stress-activated protein kinase 1 in BMP-6–induced chondrocyte differentiation

    doi: 10.1016/j.jbc.2024.107806

    Figure Lengend Snippet: I nflu ence of MSK1 deficiency on BMP-6–mediated chondrogenic differentiation. A , expression of MSK1 in ATDC5 cells. Total lysates from each transformant were prepared for Western blot analyses. The total expression levels of MSK1 and β-actin are indicated in the upper and lower panels , respectively. White and black arrowheads indicate MSK1-specific and nonspecific bands, respectively. The intensity of the band for MSK1 was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to cells carrying shControl. B , chondrogenic differentiation of ATDC5 cells with decreased expression of MSK1 in the presence of BMP-6. The cells were seeded at a high density and then stimulated with 25 ng/ml BMP-6. Seven ( upper panels ) or 14 days later ( lower panels ), the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using ImageQuant TL. The relative intensity of the stained area was calculated with respect to the cells carrying shControl with BMP-6 for 14 days. BMP, bone morphogenetic protein; MSK, mitogen- and stress-activated protein kinase; qPCR, quantitative PCR.

    Article Snippet: The intensity of each band was measured using ImageQuant TL analysis software (Cytiva) after each gel image was captured with the ImageQuant LAS 4000 mini (Amersham).

    Techniques: Expressing, Western Blot, Staining, Real-time Polymerase Chain Reaction

    R equi rement of N-terminal and C-terminal kinase activities of MSK1 for the induction of chondrocyte differentiation. A , depiction of MSK1 and its mutants used in . B , phosphorylation of MSK1 or its mutants in ATDC5 cells stimulated with BMP-6. To induce the expression of ectopic MSK1 or its mutants in ATDC5 cells, cells were cultured in the presence of tetracycline for 16 h. Thereafter, the ATDC5 transformants were stimulated with 25 ng/ml BMP-6 for 1 h. The total expression levels of phospho-MSK1 (Ser376), MSK1, phospho-CREB (Ser133), phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, and bottom panels , respectively. A black arrow and a white arrowhead indicate ectopic and endogenous phospho-MSK1 (Ser376)-specific band, respectively. A black arrowhead indicates nonspecific bands. The intensities of the bands for phospho-MSK1 (Ser376) and phospho-CREB (Ser133) were normalized to the intensities of the bands corresponding to MSK1 and β-actin, respectively. The relative intensity was calculated with respect to the 1-h treatment of cells carrying WT MSK1 with BMP-6 alone. C , chondrogenic differentiation of ATDC5 cells carrying WT MSK1 or its mutants in the presence of BMP-6. After expression of WT MSK1 or its mutants in ATDC5 cells using tetracycline as described in B , the cells were reseeded at a high density and stimulated with 25 ng/ml BMP-6. Seven ( upper photos ) or 14 days later ( lower photos ), the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using ImageQuant TL. The relative intensity of the stained area was calculated with respect to the cells carrying WT MSK1 with BMP-6 alone for 14 days. D and E , According to C , ATDC5 cells expressing WT MSK1 or MSK1 (D195A/D565A) were cultured with 25 ng/ml BMP-6 for 14 days. Thereafter, the total mRNAs from each transformant were prepared for qPCR to detect aggrecan ( D ) and collagen10a1 mRNAs ( E ). All values represent means ± SDs (n = 3). Asterisks indicate significant differences. BMP, bone morphogenetic protein; CTKD, C-terminal kinase domain; HM, hydrophobic motif; MSK, mitogen- and stress-activated protein kinase; ND, not determined; NTKD, N-terminal kinase domain; qPCR, quantitative PCR.

    Journal: The Journal of Biological Chemistry

    Article Title: Involvement of mitogen- and stress-activated protein kinase 1 in BMP-6–induced chondrocyte differentiation

    doi: 10.1016/j.jbc.2024.107806

    Figure Lengend Snippet: R equi rement of N-terminal and C-terminal kinase activities of MSK1 for the induction of chondrocyte differentiation. A , depiction of MSK1 and its mutants used in . B , phosphorylation of MSK1 or its mutants in ATDC5 cells stimulated with BMP-6. To induce the expression of ectopic MSK1 or its mutants in ATDC5 cells, cells were cultured in the presence of tetracycline for 16 h. Thereafter, the ATDC5 transformants were stimulated with 25 ng/ml BMP-6 for 1 h. The total expression levels of phospho-MSK1 (Ser376), MSK1, phospho-CREB (Ser133), phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, and bottom panels , respectively. A black arrow and a white arrowhead indicate ectopic and endogenous phospho-MSK1 (Ser376)-specific band, respectively. A black arrowhead indicates nonspecific bands. The intensities of the bands for phospho-MSK1 (Ser376) and phospho-CREB (Ser133) were normalized to the intensities of the bands corresponding to MSK1 and β-actin, respectively. The relative intensity was calculated with respect to the 1-h treatment of cells carrying WT MSK1 with BMP-6 alone. C , chondrogenic differentiation of ATDC5 cells carrying WT MSK1 or its mutants in the presence of BMP-6. After expression of WT MSK1 or its mutants in ATDC5 cells using tetracycline as described in B , the cells were reseeded at a high density and stimulated with 25 ng/ml BMP-6. Seven ( upper photos ) or 14 days later ( lower photos ), the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using ImageQuant TL. The relative intensity of the stained area was calculated with respect to the cells carrying WT MSK1 with BMP-6 alone for 14 days. D and E , According to C , ATDC5 cells expressing WT MSK1 or MSK1 (D195A/D565A) were cultured with 25 ng/ml BMP-6 for 14 days. Thereafter, the total mRNAs from each transformant were prepared for qPCR to detect aggrecan ( D ) and collagen10a1 mRNAs ( E ). All values represent means ± SDs (n = 3). Asterisks indicate significant differences. BMP, bone morphogenetic protein; CTKD, C-terminal kinase domain; HM, hydrophobic motif; MSK, mitogen- and stress-activated protein kinase; ND, not determined; NTKD, N-terminal kinase domain; qPCR, quantitative PCR.

    Article Snippet: The intensity of each band was measured using ImageQuant TL analysis software (Cytiva) after each gel image was captured with the ImageQuant LAS 4000 mini (Amersham).

    Techniques: Phospho-proteomics, Expressing, Cell Culture, Staining, Real-time Polymerase Chain Reaction

    M SK1 m utants lacking phosphorylation site targeted by p38 kinase and C-terminal kinase of MSK1. A , depiction of MSK1 and its mutants used in and . B , phosphorylation of MSK1 or its mutants in ATDC5 cells stimulated with BMP-6. After expression of WT MSK1 or its mutants in ATDC5 cells using tetracycline as described in B , the cells were stimulated with 25 ng/ml BMP-6 for 1 h and their total lysates were prepared for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, and bottom panels , respectively. A black arrow and a white arrowhead indicate ectopic and endogenous phospho-MSK1 (Ser376)-specific band, respectively. A black arrowhead indicates nonspecific bands. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells carrying WT MSK1 with BMP-6. C , comparison between serine-360 and threonine-581 in MSK1 as targeted amino acid residues for p38 kinase. After expression of WT MSK1 or its mutants in ATDC5 cells using tetracycline as described in B , the cells were stimulated with 25 ng/ml BMP-6 for 1 h and then their total lysates were prepared for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-p38 (Thr180/Tyr182), p38, phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh, eighth, and bottom panels , respectively. A black arrow and a white arrowhead indicate ectopic and endogenous phospho-MSK1 (Ser376)-specific band, respectively. A black arrowhead indicates nonspecific bands. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells carrying WT MSK1 with BMP-6. D , phosphorylation of threonine at position 581 in MSK1 upon BMP-6 stimulation. As described in B , cell lysates were prepared from each transformant. The total expression levels of phospho-MSK1 (Thr581), MSK1, phospho-Smad1/5, and β-actin are indicated in the upper, second, third, and bottom panels , respectively. The intensity of the band for phospho-MSK1 (T581) was normalized to the intensity of the band corresponding to MSK1. Relative intensity was calculated with respect to the 1-h treatment of cells carrying WT MSK1 with BMP-6. E , chondrogenic differentiation of ATDC5 cells carrying WT MSK1 or its mutants in the presence of BMP-6. After the expression of WT MSK1 or its mutants in ATDC5 cells using tetracycline as described in C , the cells were reseeded at a high density and then stimulated with 25 ng/ml BMP-6. Seven ( upper panels ) or 14 days later ( lower panels ), the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using ImageQuant TL. The relative intensity of the stained area was calculated with respect to the cells carrying WT MSK1 with BMP-6 alone for 14 days. F and G , expression of aggrecan and collagen 10a1 mRNAs in ATDC5 cells and their transformants. According to , D and E , ATDC5 cells expressing WT MSK1 or its mutants were cultured with 25 ng/ml BMP-6 for 14 days. Thereafter, total mRNAs from each transformant were prepared for qPCR to detect aggrecan ( F ) and collagen10a1 mRNAs ( G ). All values represent means ± SDs (n = 3). Asterisks indicate significant differences. BMP, bone morphogenetic protein; CREB, cAMP-response element-binding protein; MSK, mitogen- and stress-activated protein kinase; ND, not determined; qPCR, quantitative PCR.

    Journal: The Journal of Biological Chemistry

    Article Title: Involvement of mitogen- and stress-activated protein kinase 1 in BMP-6–induced chondrocyte differentiation

    doi: 10.1016/j.jbc.2024.107806

    Figure Lengend Snippet: M SK1 m utants lacking phosphorylation site targeted by p38 kinase and C-terminal kinase of MSK1. A , depiction of MSK1 and its mutants used in and . B , phosphorylation of MSK1 or its mutants in ATDC5 cells stimulated with BMP-6. After expression of WT MSK1 or its mutants in ATDC5 cells using tetracycline as described in B , the cells were stimulated with 25 ng/ml BMP-6 for 1 h and their total lysates were prepared for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, and bottom panels , respectively. A black arrow and a white arrowhead indicate ectopic and endogenous phospho-MSK1 (Ser376)-specific band, respectively. A black arrowhead indicates nonspecific bands. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells carrying WT MSK1 with BMP-6. C , comparison between serine-360 and threonine-581 in MSK1 as targeted amino acid residues for p38 kinase. After expression of WT MSK1 or its mutants in ATDC5 cells using tetracycline as described in B , the cells were stimulated with 25 ng/ml BMP-6 for 1 h and then their total lysates were prepared for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), phospho-MSK1 (Ser360), MSK1, phospho-CREB (Ser133), phospho-p38 (Thr180/Tyr182), p38, phospho-Smad1/5, Smad5, and β-actin are indicated in the upper, second, third, fourth, fifth, sixth, seventh, eighth, and bottom panels , respectively. A black arrow and a white arrowhead indicate ectopic and endogenous phospho-MSK1 (Ser376)-specific band, respectively. A black arrowhead indicates nonspecific bands. The intensity of the bands for phospho-MSK1 (Ser376) and phospho-MSK1 (Ser360) was normalized to the intensity of the band corresponding to MSK1. The intensity of the band for phospho-CREB (Ser133) was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to the 1-h treatment of cells carrying WT MSK1 with BMP-6. D , phosphorylation of threonine at position 581 in MSK1 upon BMP-6 stimulation. As described in B , cell lysates were prepared from each transformant. The total expression levels of phospho-MSK1 (Thr581), MSK1, phospho-Smad1/5, and β-actin are indicated in the upper, second, third, and bottom panels , respectively. The intensity of the band for phospho-MSK1 (T581) was normalized to the intensity of the band corresponding to MSK1. Relative intensity was calculated with respect to the 1-h treatment of cells carrying WT MSK1 with BMP-6. E , chondrogenic differentiation of ATDC5 cells carrying WT MSK1 or its mutants in the presence of BMP-6. After the expression of WT MSK1 or its mutants in ATDC5 cells using tetracycline as described in C , the cells were reseeded at a high density and then stimulated with 25 ng/ml BMP-6. Seven ( upper panels ) or 14 days later ( lower panels ), the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using ImageQuant TL. The relative intensity of the stained area was calculated with respect to the cells carrying WT MSK1 with BMP-6 alone for 14 days. F and G , expression of aggrecan and collagen 10a1 mRNAs in ATDC5 cells and their transformants. According to , D and E , ATDC5 cells expressing WT MSK1 or its mutants were cultured with 25 ng/ml BMP-6 for 14 days. Thereafter, total mRNAs from each transformant were prepared for qPCR to detect aggrecan ( F ) and collagen10a1 mRNAs ( G ). All values represent means ± SDs (n = 3). Asterisks indicate significant differences. BMP, bone morphogenetic protein; CREB, cAMP-response element-binding protein; MSK, mitogen- and stress-activated protein kinase; ND, not determined; qPCR, quantitative PCR.

    Article Snippet: The intensity of each band was measured using ImageQuant TL analysis software (Cytiva) after each gel image was captured with the ImageQuant LAS 4000 mini (Amersham).

    Techniques: Phospho-proteomics, Expressing, Western Blot, Comparison, Staining, Cell Culture, Binding Assay, Real-time Polymerase Chain Reaction

    Req u irement of autophosphorylation of Ser 376 in MSK1 for its kinase activity upon BMP-6 stimulation. A , chondrogenic differentiation of ATDC5 cells carrying WT MSK1 or MSK1 (S376A) mutant in the presence of BMP-6. After the expression of WT MSK1 or MSK1 (S376A) mutant in ATDC5 cells using tetracycline as described in C , the cells were reseeded at a high density and then stimulated with 25 ng/ml BMP-6. Seven ( upper panels ) or 14 days later ( lower panels ), the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using ImageQuant TL. The relative intensity of the stained area was calculated with respect to the cells carrying WT MSK1 with BMP-6 for 14 days. B and C , expression of aggrecan and collagen10a1 mRNAs in ATDC5 cells and their transformant. According to , D and E , ATDC5 cells expressing the WT MSK1 or its mutant were cultured with 25 ng/ml BMP-6 for 14 days. Thereafter, total mRNAs from each transformant were prepared for qPCR to detect aggrecan ( B ) and collagen10a1 mRNAs ( C ). All values represent means ± SDs (n = 3). Asterisks indicate significant differences. BMP, bone morphogenetic protein; MSK, mitogen- and stress-activated protein kinase; qPCR, quantitative PCR.

    Journal: The Journal of Biological Chemistry

    Article Title: Involvement of mitogen- and stress-activated protein kinase 1 in BMP-6–induced chondrocyte differentiation

    doi: 10.1016/j.jbc.2024.107806

    Figure Lengend Snippet: Req u irement of autophosphorylation of Ser 376 in MSK1 for its kinase activity upon BMP-6 stimulation. A , chondrogenic differentiation of ATDC5 cells carrying WT MSK1 or MSK1 (S376A) mutant in the presence of BMP-6. After the expression of WT MSK1 or MSK1 (S376A) mutant in ATDC5 cells using tetracycline as described in C , the cells were reseeded at a high density and then stimulated with 25 ng/ml BMP-6. Seven ( upper panels ) or 14 days later ( lower panels ), the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using ImageQuant TL. The relative intensity of the stained area was calculated with respect to the cells carrying WT MSK1 with BMP-6 for 14 days. B and C , expression of aggrecan and collagen10a1 mRNAs in ATDC5 cells and their transformant. According to , D and E , ATDC5 cells expressing the WT MSK1 or its mutant were cultured with 25 ng/ml BMP-6 for 14 days. Thereafter, total mRNAs from each transformant were prepared for qPCR to detect aggrecan ( B ) and collagen10a1 mRNAs ( C ). All values represent means ± SDs (n = 3). Asterisks indicate significant differences. BMP, bone morphogenetic protein; MSK, mitogen- and stress-activated protein kinase; qPCR, quantitative PCR.

    Article Snippet: The intensity of each band was measured using ImageQuant TL analysis software (Cytiva) after each gel image was captured with the ImageQuant LAS 4000 mini (Amersham).

    Techniques: Activity Assay, Mutagenesis, Expressing, Staining, Cell Culture, Real-time Polymerase Chain Reaction

    Effect of the BMP/Smad-dependent pathway on BMP-6–mediated chondrogenic differentiation. A , expression of Smad4 in ATDC5 cells. Total lysates from each transformant were prepared for Western blot analyses. The total expression levels of Smad4 and β-actin are indicated in the upper and lower panels , respectively. The intensity of the band for Smad4 was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to cells carrying shControl. B , chondrogenic differentiation of ATDC5 cells with decreased expression of Smad4 in the presence of BMP-6. The cells were seeded at a high density and then stimulated with 25 ng/ml BMP-6. Seven ( upper panels ) or 14 days later ( lower panels ), the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using ImageQuant TL. The relative intensity of the stained area was calculated with respect to the cells carrying shControl with BMP-6 for 14 days. C and D , expression of aggrecan and collagen10a1 mRNA in ATDC5 cells with Smad4 knockdown. According to <xref ref-type=Fig. 2 , D and E , ATDC5 cells with decreased expression of Smad4 were cultured with 25 ng/ml BMP-6 for 14 days. Thereafter, total mRNAs from each transformant were prepared for qPCR to detect aggrecan ( C ) and collagen10a1 mRNAs ( D ). All values represent means ± SDs (n = 3). Asterisks indicate significant differences. E , effect of Smad4 deficiency on BMP-6–induced p38 phosphorylation. The cells were stimulated with 25 ng/ml BMP-6 for 1 h and then their total lysates were prepared for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), MSK1, phospho-p38 (Thr180/Tyr182), phospho-Smad1/5, and β-actin are indicated in the upper, second, third, fourth, and bottom panels , respectively. The intensity of the band for phospho-MSK1 (S376) was normalized to the intensity of the band corresponding to MSK1. Relative intensity was calculated with respect to the 1-h treatment of cells carrying shControl with BMP-6. BMP, bone morphogenetic protein; MSK, mitogen- and stress-activated protein kinase; ND, not determined; qPCR, quantitative PCR. " width="100%" height="100%">

    Journal: The Journal of Biological Chemistry

    Article Title: Involvement of mitogen- and stress-activated protein kinase 1 in BMP-6–induced chondrocyte differentiation

    doi: 10.1016/j.jbc.2024.107806

    Figure Lengend Snippet: Effect of the BMP/Smad-dependent pathway on BMP-6–mediated chondrogenic differentiation. A , expression of Smad4 in ATDC5 cells. Total lysates from each transformant were prepared for Western blot analyses. The total expression levels of Smad4 and β-actin are indicated in the upper and lower panels , respectively. The intensity of the band for Smad4 was normalized to the intensity of the band corresponding to β-actin. Relative intensity was calculated with respect to cells carrying shControl. B , chondrogenic differentiation of ATDC5 cells with decreased expression of Smad4 in the presence of BMP-6. The cells were seeded at a high density and then stimulated with 25 ng/ml BMP-6. Seven ( upper panels ) or 14 days later ( lower panels ), the cells were stained with Alcian blue. Representative images are shown. The intensity of the area stained with Alcian blue was measured using ImageQuant TL. The relative intensity of the stained area was calculated with respect to the cells carrying shControl with BMP-6 for 14 days. C and D , expression of aggrecan and collagen10a1 mRNA in ATDC5 cells with Smad4 knockdown. According to Fig. 2 , D and E , ATDC5 cells with decreased expression of Smad4 were cultured with 25 ng/ml BMP-6 for 14 days. Thereafter, total mRNAs from each transformant were prepared for qPCR to detect aggrecan ( C ) and collagen10a1 mRNAs ( D ). All values represent means ± SDs (n = 3). Asterisks indicate significant differences. E , effect of Smad4 deficiency on BMP-6–induced p38 phosphorylation. The cells were stimulated with 25 ng/ml BMP-6 for 1 h and then their total lysates were prepared for Western blot analyses. The total expression levels of phospho-MSK1 (Ser376), MSK1, phospho-p38 (Thr180/Tyr182), phospho-Smad1/5, and β-actin are indicated in the upper, second, third, fourth, and bottom panels , respectively. The intensity of the band for phospho-MSK1 (S376) was normalized to the intensity of the band corresponding to MSK1. Relative intensity was calculated with respect to the 1-h treatment of cells carrying shControl with BMP-6. BMP, bone morphogenetic protein; MSK, mitogen- and stress-activated protein kinase; ND, not determined; qPCR, quantitative PCR.

    Article Snippet: The intensity of each band was measured using ImageQuant TL analysis software (Cytiva) after each gel image was captured with the ImageQuant LAS 4000 mini (Amersham).

    Techniques: Expressing, Western Blot, Staining, Knockdown, Cell Culture, Phospho-proteomics, Real-time Polymerase Chain Reaction