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blue alkaline phosphatase alp substrate kit  (Vector Laboratories)


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    Structured Review

    Vector Laboratories blue alkaline phosphatase alp substrate kit
    Maternal exercise alters growth plate morphometry and bone neovascularization. (A) Fluorescent staining for collagen type 10 alpha 1 chain (Col10a1) in the proximal hypertrophic zone (HZ) of embryonic day 17.5 (E17.5) humeri. Scale bar = 500 μm. (B) HZ length. (C) Fluorescent staining for alkaline <t>phosphatase</t> <t>(ALP)</t> activity in E17.5 humeri. Scale bar = 250 μm. (D) Primary ossification center (POC) ALP activity and (E) bone collar (BC) ALP activity. (F) Fluorescent staining for blood vessels (endomucin) in E17.5 humeri. Scale bar = 250 μm. (G) Medullary vessel area, (H) Medullary vessel density, (I) BC vessel area, (J) BC vessel density, (K) metaphyseal vessel area, and (L) metaphyseal vessel density. Fetuses from the same litter are marked by the same color datapoint. * p < 0.05 using a Student's t ‐test.
    Blue Alkaline Phosphatase Alp Substrate Kit, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 96/100, based on 1591 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/blue+alkaline+phosphatase+alp+substrate+kit/Vector+Blue+Alkaline+Phosphatase+(Blue+AP)+Substrate+Kit/pmc12701528-120-9-16
    Average 96 stars, based on 1591 article reviews
    blue alkaline phosphatase alp substrate kit - by Bioz Stars, 2026-10
    96/100 stars

    Images

    1) Product Images from "Maternal Exercise Rescues Fetal Akinesia‐Impaired Joint and Bone Development"

    Article Title: Maternal Exercise Rescues Fetal Akinesia‐Impaired Joint and Bone Development

    Journal: The FASEB Journal

    doi: 10.1096/fj.202503192R

    Maternal exercise alters growth plate morphometry and bone neovascularization. (A) Fluorescent staining for collagen type 10 alpha 1 chain (Col10a1) in the proximal hypertrophic zone (HZ) of embryonic day 17.5 (E17.5) humeri. Scale bar = 500 μm. (B) HZ length. (C) Fluorescent staining for alkaline phosphatase (ALP) activity in E17.5 humeri. Scale bar = 250 μm. (D) Primary ossification center (POC) ALP activity and (E) bone collar (BC) ALP activity. (F) Fluorescent staining for blood vessels (endomucin) in E17.5 humeri. Scale bar = 250 μm. (G) Medullary vessel area, (H) Medullary vessel density, (I) BC vessel area, (J) BC vessel density, (K) metaphyseal vessel area, and (L) metaphyseal vessel density. Fetuses from the same litter are marked by the same color datapoint. * p < 0.05 using a Student's t ‐test.
    Figure Legend Snippet: Maternal exercise alters growth plate morphometry and bone neovascularization. (A) Fluorescent staining for collagen type 10 alpha 1 chain (Col10a1) in the proximal hypertrophic zone (HZ) of embryonic day 17.5 (E17.5) humeri. Scale bar = 500 μm. (B) HZ length. (C) Fluorescent staining for alkaline phosphatase (ALP) activity in E17.5 humeri. Scale bar = 250 μm. (D) Primary ossification center (POC) ALP activity and (E) bone collar (BC) ALP activity. (F) Fluorescent staining for blood vessels (endomucin) in E17.5 humeri. Scale bar = 250 μm. (G) Medullary vessel area, (H) Medullary vessel density, (I) BC vessel area, (J) BC vessel density, (K) metaphyseal vessel area, and (L) metaphyseal vessel density. Fetuses from the same litter are marked by the same color datapoint. * p < 0.05 using a Student's t ‐test.

    Techniques Used: Staining, Activity Assay

    Related Articles

    Staining:

    Article Title: Maternal Exercise Rescues Fetal Akinesia‐Impaired Joint and Bone Development
    Article Snippet: For all protocols, samples were mounted using Fluoromount‐GT (Thermo Fisher Scientific), imaged using the ZEISS AxioScan.Z1 Slide Scanner (ZEISS, Oberkochen, Germany), and analyzed using ImageJ/Fiji. .. Alkaline phosphatase (ALP) staining was conducted using the Vector blue Alkaline Phosphatase (ALP) substrate kit (SK‐5300, Vector Laboratories, Newark, CA, USA) according to the manufacturer's instructions. .. Phalloidin staining with Hoechst counterstain (Thermo Fisher Scientific) was conducted using Alexa Fluor 647 Phalloidin (A22287, Thermo Fisher Scientific) according to the manufacturer's instructions.

    Article Title: Multisite studies for validation and improvement of a highly efficient culture assay for detection of undifferentiated human pluripotent stem cells intermingled in cell therapy products.
    Article Snippet: Each 1 mL of cell suspension using E8F +Y medium or AK03N+Y medium was respectively seeded on 10-cm culture dishes coated with laminin-521 (BioLamina AB) [23] or 10- cm culture dishes containing iMatrix-511 solution (0.25 mg/cm2) [25]. .. The day after the seeding, media were replaced with each respective medium without Y-27632, and after 5 or 6 days of incubation at 37°C in 5% CO2, the dishes were fixed with 4% paraformaldehyde phosphate buffer solution and stained using a Vector Blue Alkaline Phosphatase (ALP) substrate kit (Vector Laboratories, Inc) according to the manufacturer’s instructions. ..

    Article Title: International evaluation study of a highly efficient culture assay for detection of residual human pluripotent stem cells in cell therapies.
    Article Snippet: Four wells were prepared for each concentration, and wells in which only 1 × 106 iCell cardiomyocytes were seeded (nonspiked sample) were prepared for each experiment (n = 1 or 2). .. Three days after seeding, the media were replaced with Essential 8 Flex medium, and after a total of 7 days of incubation at 37◦C in 5% CO2, the wells were fixed and stained using a Vector Blue alkaline phosphatase (ALP) substrate kit (Vector Laboratories, Inc.) according to a previously reported method [14]. ..

    Article Title: Maternal exercise rescues fetal akinesia-impaired joint and bone development
    Article Snippet: For all protocols, samples were mounted using Fluoromount-GT (Thermo Fisher Scientific), imaged using the ZEISS AxioScan.Z1 Slide Scanner (ZEISS, Oberkochen, Germany), and analyzed using ImageJ/Fiji. .. Alkaline phosphatase (ALP) staining was conducted using the Vector blue Alkaline Phosphatase (ALP) substrate kit (SK-5300, Vector Laboratories, Newark, CA, USA) according to the manufacturer’s instructions. .. Phalloidin staining with Hoechst counterstain (Thermo Fisher Scientific) was conducted using Alexa Fluor 647 Phalloidin (A22287, Thermo Fisher Scientific) according to manufacturer’s instructions.

    Incubation:

    Article Title: Multisite studies for validation and improvement of a highly efficient culture assay for detection of undifferentiated human pluripotent stem cells intermingled in cell therapy products.
    Article Snippet: Each 1 mL of cell suspension using E8F +Y medium or AK03N+Y medium was respectively seeded on 10-cm culture dishes coated with laminin-521 (BioLamina AB) [23] or 10- cm culture dishes containing iMatrix-511 solution (0.25 mg/cm2) [25]. .. The day after the seeding, media were replaced with each respective medium without Y-27632, and after 5 or 6 days of incubation at 37°C in 5% CO2, the dishes were fixed with 4% paraformaldehyde phosphate buffer solution and stained using a Vector Blue Alkaline Phosphatase (ALP) substrate kit (Vector Laboratories, Inc) according to the manufacturer’s instructions. ..

    Article Title: International evaluation study of a highly efficient culture assay for detection of residual human pluripotent stem cells in cell therapies.
    Article Snippet: Four wells were prepared for each concentration, and wells in which only 1 × 106 iCell cardiomyocytes were seeded (nonspiked sample) were prepared for each experiment (n = 1 or 2). .. Three days after seeding, the media were replaced with Essential 8 Flex medium, and after a total of 7 days of incubation at 37◦C in 5% CO2, the wells were fixed and stained using a Vector Blue alkaline phosphatase (ALP) substrate kit (Vector Laboratories, Inc.) according to a previously reported method [14]. ..



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    Vector Laboratories blue alkaline phosphatase alp substrate kit
    Maternal exercise alters growth plate morphometry and bone neovascularization. (A) Fluorescent staining for collagen type 10 alpha 1 chain (Col10a1) in the proximal hypertrophic zone (HZ) of embryonic day 17.5 (E17.5) humeri. Scale bar = 500 μm. (B) HZ length. (C) Fluorescent staining for alkaline <t>phosphatase</t> <t>(ALP)</t> activity in E17.5 humeri. Scale bar = 250 μm. (D) Primary ossification center (POC) ALP activity and (E) bone collar (BC) ALP activity. (F) Fluorescent staining for blood vessels (endomucin) in E17.5 humeri. Scale bar = 250 μm. (G) Medullary vessel area, (H) Medullary vessel density, (I) BC vessel area, (J) BC vessel density, (K) metaphyseal vessel area, and (L) metaphyseal vessel density. Fetuses from the same litter are marked by the same color datapoint. * p < 0.05 using a Student's t ‐test.
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    https://www.bioz.com/product/blue+alkaline+phosphatase+alp+substrate+kit/Vector+Blue+Alkaline+Phosphatase+(Blue+AP)+Substrate+Kit/pmc12701528-120-9-16
    Average 96 stars, based on 1 article reviews
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    Vector Laboratories vector blue alkaline phosphatase alp substrate kit
    Maternal exercise alters growth plate morphometry and bone neovascularization. (A) Fluorescent staining for collagen type 10 alpha 1 chain (Col10a1) in the proximal hypertrophic zone (HZ) of embryonic day 17.5 (E17.5) humeri. Scale bar = 500 μm. (B) HZ length. (C) Fluorescent staining for alkaline <t>phosphatase</t> <t>(ALP)</t> activity in E17.5 humeri. Scale bar = 250 μm. (D) Primary ossification center (POC) ALP activity and (E) bone collar (BC) ALP activity. (F) Fluorescent staining for blood vessels (endomucin) in E17.5 humeri. Scale bar = 250 μm. (G) Medullary vessel area, (H) Medullary vessel density, (I) BC vessel area, (J) BC vessel density, (K) metaphyseal vessel area, and (L) metaphyseal vessel density. Fetuses from the same litter are marked by the same color datapoint. * p < 0.05 using a Student's t ‐test.
    Vector Blue Alkaline Phosphatase Alp Substrate Kit, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/blue+alkaline+phosphatase+alp+substrate+kit/Vector+Blue+Alkaline+Phosphatase+(Blue+AP)+Substrate+Kit/pmc11231703-61-45-52
    Average 96 stars, based on 1 article reviews
    vector blue alkaline phosphatase alp substrate kit - by Bioz Stars, 2026-10
    96/100 stars
      Buy from Supplier

    96
    Vector Laboratories alp substrate kit
    Maternal exercise alters growth plate morphometry and bone neovascularization. (A) Fluorescent staining for collagen type 10 alpha 1 chain (Col10a1) in the proximal hypertrophic zone (HZ) of embryonic day 17.5 (E17.5) humeri. Scale bar = 500 μm. (B) HZ length. (C) Fluorescent staining for alkaline <t>phosphatase</t> <t>(ALP)</t> activity in E17.5 humeri. Scale bar = 250 μm. (D) Primary ossification center (POC) ALP activity and (E) bone collar (BC) ALP activity. (F) Fluorescent staining for blood vessels (endomucin) in E17.5 humeri. Scale bar = 250 μm. (G) Medullary vessel area, (H) Medullary vessel density, (I) BC vessel area, (J) BC vessel density, (K) metaphyseal vessel area, and (L) metaphyseal vessel density. Fetuses from the same litter are marked by the same color datapoint. * p < 0.05 using a Student's t ‐test.
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    https://www.bioz.com/product/blue+alkaline+phosphatase+alp+substrate+kit/Vector+Blue+Alkaline+Phosphatase+(Blue+AP)+Substrate+Kit/pmc06145045-87-24-27
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    Vector Laboratories alkaline phosphatase alp staining kit
    Lipid scarcity suppressed bone marrow mesenchymal stromal cell (BMSC) osteogenesis and Wnt/beta-catenin signaling activation. (A) Alkaline phosphatase <t>(ALP)</t> staining of BMSCs 7 days <t>after</t> <t>osteogenic</t> induction in culture medium with normal fetal bovine serum (FBS; 10%), lipid-reduced serum (LRS; 10%), SD, and LRS (10%) with 75 μM oleate treatment. (B) ALP activity of BMSCs 5, 7, and 11 days after osteogenic induction in different culture mediums. (C) Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assays revealed significantly decreased cell viability in SD treatment BMSCs compared to FBS treatment group, while the cell viability was not significantly changed in LRS and LRS + OL group. (D,E) Western blots showed that the protein expression levels of osteogenic markers Type I Collagen (COL1) and RUNX Family Transcription Factor 2 (RUNX2) and beta-catenin were significantly decreased after lipid deprivation and partially restored by additional oleate treatment 7 days after induction. (F) qRT-PCR revealed that the mRNA expression levels of osteogenic genes, Alpl , Sp7 , and Col1a1 , as well as Wnt target genes Tcf1 , Lef1 , and Axin2 , were significantly decreased after lipid deprivation and partially restored by additional oleate treatment 7 days after induction, * p < 0.05, ** p < 0.01, and *** p < 0.001.
    Alkaline Phosphatase Alp Staining Kit, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Maternal exercise alters growth plate morphometry and bone neovascularization. (A) Fluorescent staining for collagen type 10 alpha 1 chain (Col10a1) in the proximal hypertrophic zone (HZ) of embryonic day 17.5 (E17.5) humeri. Scale bar = 500 μm. (B) HZ length. (C) Fluorescent staining for alkaline phosphatase (ALP) activity in E17.5 humeri. Scale bar = 250 μm. (D) Primary ossification center (POC) ALP activity and (E) bone collar (BC) ALP activity. (F) Fluorescent staining for blood vessels (endomucin) in E17.5 humeri. Scale bar = 250 μm. (G) Medullary vessel area, (H) Medullary vessel density, (I) BC vessel area, (J) BC vessel density, (K) metaphyseal vessel area, and (L) metaphyseal vessel density. Fetuses from the same litter are marked by the same color datapoint. * p < 0.05 using a Student's t ‐test.

    Journal: The FASEB Journal

    Article Title: Maternal Exercise Rescues Fetal Akinesia‐Impaired Joint and Bone Development

    doi: 10.1096/fj.202503192R

    Figure Lengend Snippet: Maternal exercise alters growth plate morphometry and bone neovascularization. (A) Fluorescent staining for collagen type 10 alpha 1 chain (Col10a1) in the proximal hypertrophic zone (HZ) of embryonic day 17.5 (E17.5) humeri. Scale bar = 500 μm. (B) HZ length. (C) Fluorescent staining for alkaline phosphatase (ALP) activity in E17.5 humeri. Scale bar = 250 μm. (D) Primary ossification center (POC) ALP activity and (E) bone collar (BC) ALP activity. (F) Fluorescent staining for blood vessels (endomucin) in E17.5 humeri. Scale bar = 250 μm. (G) Medullary vessel area, (H) Medullary vessel density, (I) BC vessel area, (J) BC vessel density, (K) metaphyseal vessel area, and (L) metaphyseal vessel density. Fetuses from the same litter are marked by the same color datapoint. * p < 0.05 using a Student's t ‐test.

    Article Snippet: Alkaline phosphatase (ALP) staining was conducted using the Vector blue Alkaline Phosphatase (ALP) substrate kit (SK‐5300, Vector Laboratories, Newark, CA, USA) according to the manufacturer's instructions.

    Techniques: Staining, Activity Assay

    Lipid scarcity suppressed bone marrow mesenchymal stromal cell (BMSC) osteogenesis and Wnt/beta-catenin signaling activation. (A) Alkaline phosphatase (ALP) staining of BMSCs 7 days after osteogenic induction in culture medium with normal fetal bovine serum (FBS; 10%), lipid-reduced serum (LRS; 10%), SD, and LRS (10%) with 75 μM oleate treatment. (B) ALP activity of BMSCs 5, 7, and 11 days after osteogenic induction in different culture mediums. (C) Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assays revealed significantly decreased cell viability in SD treatment BMSCs compared to FBS treatment group, while the cell viability was not significantly changed in LRS and LRS + OL group. (D,E) Western blots showed that the protein expression levels of osteogenic markers Type I Collagen (COL1) and RUNX Family Transcription Factor 2 (RUNX2) and beta-catenin were significantly decreased after lipid deprivation and partially restored by additional oleate treatment 7 days after induction. (F) qRT-PCR revealed that the mRNA expression levels of osteogenic genes, Alpl , Sp7 , and Col1a1 , as well as Wnt target genes Tcf1 , Lef1 , and Axin2 , were significantly decreased after lipid deprivation and partially restored by additional oleate treatment 7 days after induction, * p < 0.05, ** p < 0.01, and *** p < 0.001.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: LRP5-Mediated Lipid Uptake Modulates Osteogenic Differentiation of Bone Marrow Mesenchymal Stromal Cells

    doi: 10.3389/fcell.2021.766815

    Figure Lengend Snippet: Lipid scarcity suppressed bone marrow mesenchymal stromal cell (BMSC) osteogenesis and Wnt/beta-catenin signaling activation. (A) Alkaline phosphatase (ALP) staining of BMSCs 7 days after osteogenic induction in culture medium with normal fetal bovine serum (FBS; 10%), lipid-reduced serum (LRS; 10%), SD, and LRS (10%) with 75 μM oleate treatment. (B) ALP activity of BMSCs 5, 7, and 11 days after osteogenic induction in different culture mediums. (C) Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assays revealed significantly decreased cell viability in SD treatment BMSCs compared to FBS treatment group, while the cell viability was not significantly changed in LRS and LRS + OL group. (D,E) Western blots showed that the protein expression levels of osteogenic markers Type I Collagen (COL1) and RUNX Family Transcription Factor 2 (RUNX2) and beta-catenin were significantly decreased after lipid deprivation and partially restored by additional oleate treatment 7 days after induction. (F) qRT-PCR revealed that the mRNA expression levels of osteogenic genes, Alpl , Sp7 , and Col1a1 , as well as Wnt target genes Tcf1 , Lef1 , and Axin2 , were significantly decreased after lipid deprivation and partially restored by additional oleate treatment 7 days after induction, * p < 0.05, ** p < 0.01, and *** p < 0.001.

    Article Snippet: After 5/7/11 days of osteogenic induction, the osteogenic differentiation was assessed by alkaline phosphatase (ALP) staining kit (VectorLab) in the 12-well plate.

    Techniques: Activation Assay, Staining, Activity Assay, Western Blot, Expressing, Quantitative RT-PCR

    The suppressed osteogenesis was rescued by oleate supplementation in a dosage-dependent manner. (A) Alkaline phosphatase (ALP) staining of bone marrow mesenchymal stromal cells (BMSCs) 7 days after osteogenic induction in culture medium with normal fetal bovine serum (FBS; 10%), lipid-reduced serum (LRS; 10%), and LRS (10%) with increasing dosages of oleate (25, 50, 75, 100 μM). (B) ALP activity of BMSCs 5, 7, and 11 days after osteogenic induction in different culture mediums. (C,D) Western blots showed that the suppressed protein expression levels of osteogenic markers COL1 and RUNX2 and beta-catenin by lipid deprivation were restored by additional oleate treatment in a dosage-dependent manner. (E) qRT-PCR revealed that the suppressed mRNA expression levels of osteogenic genes Alpl , Sp7 , and Col1a1 and Wnt target genes Tcf1 , Lef1 , and Axin2 by lipid deprivation were restored by additional oleate treatment in a dosage-dependent manner, * p < 0.05, ** p < 0.01, and *** p < 0.001.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: LRP5-Mediated Lipid Uptake Modulates Osteogenic Differentiation of Bone Marrow Mesenchymal Stromal Cells

    doi: 10.3389/fcell.2021.766815

    Figure Lengend Snippet: The suppressed osteogenesis was rescued by oleate supplementation in a dosage-dependent manner. (A) Alkaline phosphatase (ALP) staining of bone marrow mesenchymal stromal cells (BMSCs) 7 days after osteogenic induction in culture medium with normal fetal bovine serum (FBS; 10%), lipid-reduced serum (LRS; 10%), and LRS (10%) with increasing dosages of oleate (25, 50, 75, 100 μM). (B) ALP activity of BMSCs 5, 7, and 11 days after osteogenic induction in different culture mediums. (C,D) Western blots showed that the suppressed protein expression levels of osteogenic markers COL1 and RUNX2 and beta-catenin by lipid deprivation were restored by additional oleate treatment in a dosage-dependent manner. (E) qRT-PCR revealed that the suppressed mRNA expression levels of osteogenic genes Alpl , Sp7 , and Col1a1 and Wnt target genes Tcf1 , Lef1 , and Axin2 by lipid deprivation were restored by additional oleate treatment in a dosage-dependent manner, * p < 0.05, ** p < 0.01, and *** p < 0.001.

    Article Snippet: After 5/7/11 days of osteogenic induction, the osteogenic differentiation was assessed by alkaline phosphatase (ALP) staining kit (VectorLab) in the 12-well plate.

    Techniques: Staining, Activity Assay, Western Blot, Expressing, Quantitative RT-PCR

    Low-density lipoprotein receptor-related protein 5 (Lrp5 ) ablation in bone marrow mesenchymal stromal cells (BMSCs) impairs lipid uptake and osteogenesis. (A) BODIPY TM staining revealed significantly decreased lipid droplets in BMSC cytoplasm after Lrp5 ablation using adenoviral Cre-recombinase. (B) Quantified fluorescence intensity of lipid droplets in BMSC cytoplasm indicating impaired lipid uptake of Lrp5 -KO BMSCs. (C) LRP5 protein expression in BMSCs harvested from postnatal day 0 (P0) control mice, conditional knockout (CKO) mice, Lrp5 -control BMSC, and Lrp5 -KO BMSCs. (D) qRT-PCR validated the knockout effect of adenoviral-Cre infection on Lrp5 fl/fl BMSCs. (E) Alkaline phosphatase (ALP) staining of control and Lrp5 -KO BMSCs 7 days after osteogenic induction in culture medium with normal fetal bovine serum (FBS), lipid-reduced serum (LRS), LRS with 100 μM oleate treatment, and LRS with recombinant 100 ng/ml Wnt3a treatment, respectively. (F) ALP activity of BMSCs 7 days after osteogenic induction in different culture mediums. (G) mRNA expression levels in control and Lrp5 -KO BMSCs of osteogenic genes Alpl , Sp7 , and Col1a1 and Wnt target genes Tcf1 , Lef1 , and Axin2 after 7 days of osteogenic induction in FBS or LRS culture medium. (H) mRNA expression levels in control and Lrp5 -KO BMSCs of osteogenic genes Alpl , Sp7 , and Col1a1 as well as Wnt target genes Tcf1 , Lef1 , and Axin2 after 7 days of induction in LRS culture medium with or without Wnt3a treatment, * p < 0.05, ** p < 0.01, and *** p < 0.001.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: LRP5-Mediated Lipid Uptake Modulates Osteogenic Differentiation of Bone Marrow Mesenchymal Stromal Cells

    doi: 10.3389/fcell.2021.766815

    Figure Lengend Snippet: Low-density lipoprotein receptor-related protein 5 (Lrp5 ) ablation in bone marrow mesenchymal stromal cells (BMSCs) impairs lipid uptake and osteogenesis. (A) BODIPY TM staining revealed significantly decreased lipid droplets in BMSC cytoplasm after Lrp5 ablation using adenoviral Cre-recombinase. (B) Quantified fluorescence intensity of lipid droplets in BMSC cytoplasm indicating impaired lipid uptake of Lrp5 -KO BMSCs. (C) LRP5 protein expression in BMSCs harvested from postnatal day 0 (P0) control mice, conditional knockout (CKO) mice, Lrp5 -control BMSC, and Lrp5 -KO BMSCs. (D) qRT-PCR validated the knockout effect of adenoviral-Cre infection on Lrp5 fl/fl BMSCs. (E) Alkaline phosphatase (ALP) staining of control and Lrp5 -KO BMSCs 7 days after osteogenic induction in culture medium with normal fetal bovine serum (FBS), lipid-reduced serum (LRS), LRS with 100 μM oleate treatment, and LRS with recombinant 100 ng/ml Wnt3a treatment, respectively. (F) ALP activity of BMSCs 7 days after osteogenic induction in different culture mediums. (G) mRNA expression levels in control and Lrp5 -KO BMSCs of osteogenic genes Alpl , Sp7 , and Col1a1 and Wnt target genes Tcf1 , Lef1 , and Axin2 after 7 days of osteogenic induction in FBS or LRS culture medium. (H) mRNA expression levels in control and Lrp5 -KO BMSCs of osteogenic genes Alpl , Sp7 , and Col1a1 as well as Wnt target genes Tcf1 , Lef1 , and Axin2 after 7 days of induction in LRS culture medium with or without Wnt3a treatment, * p < 0.05, ** p < 0.01, and *** p < 0.001.

    Article Snippet: After 5/7/11 days of osteogenic induction, the osteogenic differentiation was assessed by alkaline phosphatase (ALP) staining kit (VectorLab) in the 12-well plate.

    Techniques: Staining, Fluorescence, Expressing, Knock-Out, Quantitative RT-PCR, Infection, Recombinant, Activity Assay