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Abmart Inc phospho runx2
Phospho Runx2, supplied by Abmart 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/phospho+runx2/anti+proteins+sdf2+target/pm42120379-461-35-38
Average 86 stars, based on 1 article reviews

phospho runx2 - by Bioz Stars, 2026-09

86/100 stars

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Membrane:

Article Title: Tumor-derived GDF15 induces CCN3⁺ Schwann cells to promote cancer pain in pancreatic cancer.
Article Snippet: .. The eluted proteins were resolved by SDS-PAGE, transferred to a PVDF membrane, blocked with 5% BSA for 1 hour at room temperature, followed by an overnight incubation at 4°C with the following respective primary antibodies: Phospho-RUNX2 (1:1000; TA7379, Abmart), AKT (1:1000; T55561, Abmart), GAPDH (1:5000; 60004-1-Ig, Proteintech) and detected using HRP-conjugated secondary antibodies and chemiluminescence. ..

Incubation:

Image Search Results

PMSC differentiation under low oxygen tension and its effect on osteogenic differentiation and multipotency. PMSCs were cultured for 14 days in nondifferentiation or osteogenic differentiation conditions containing 15% FBS in room air (20% O 2 ) or low oxygen levels (1% O 2 ). Treatments were stopped after 1, 3, 7, 10, and 14 days for alizarin red staining to confirm (a) PMSC differentiation morphology and quantified in (b) (two-way ANOVA, P < 0.05, N = 4). ∗ indicates significance between room air and low oxygen tension at each time point; lowercase letter (a, b, and c) indicates significance between time points within room air condition, uppercase letter (A, B) indicates within low oxygen tension. (c) Immunoblots showing protein levels of pluripotency-associated and differentiation markers from cell lysates isolated at 3, 7, and 14 days. Quantifications of day 14 samples show protein levels for (d) OCT4, (e) SOX2, (f) RUNX2, and (g) OPN in nondifferentiation and differentiation conditions. Quantification levels shown were normalized to β -actin, a protein loading control (two-way ANOVA, P < 0.05, N = 3). ∗ indicates significance between room air and low oxygen tension; # indicates significance between nondifferentiation and differentiation within the same oxygen tension.

Journal: Stem Cells International

Article Title: Regulation of Osteogenic Differentiation of Placental-Derived Mesenchymal Stem Cells by Insulin-Like Growth Factors and Low Oxygen Tension

doi: 10.1155/2017/4576327

Figure Lengend Snippet: PMSC differentiation under low oxygen tension and its effect on osteogenic differentiation and multipotency. PMSCs were cultured for 14 days in nondifferentiation or osteogenic differentiation conditions containing 15% FBS in room air (20% O 2 ) or low oxygen levels (1% O 2 ). Treatments were stopped after 1, 3, 7, 10, and 14 days for alizarin red staining to confirm (a) PMSC differentiation morphology and quantified in (b) (two-way ANOVA, P < 0.05, N = 4). ∗ indicates significance between room air and low oxygen tension at each time point; lowercase letter (a, b, and c) indicates significance between time points within room air condition, uppercase letter (A, B) indicates within low oxygen tension. (c) Immunoblots showing protein levels of pluripotency-associated and differentiation markers from cell lysates isolated at 3, 7, and 14 days. Quantifications of day 14 samples show protein levels for (d) OCT4, (e) SOX2, (f) RUNX2, and (g) OPN in nondifferentiation and differentiation conditions. Quantification levels shown were normalized to β -actin, a protein loading control (two-way ANOVA, P < 0.05, N = 3). ∗ indicates significance between room air and low oxygen tension; # indicates significance between nondifferentiation and differentiation within the same oxygen tension.

Article Snippet: For the osteogenic differentiation markers, we used RUNX2 (#8486) (Cell Signaling Technologies, Burlington, ON), phospho-RUNX2 (PA5-12988) (Thermo Fisher Scientific, Burlington, ON), and OPN (K-20, sc-1059) (Santa Cruz Biotech., Santa Cruz, CA).

Techniques: Cell Culture, Staining, Western Blot, Isolation

Effect of low oxygen tension preconditioning on PMSC osteogenic differentiation and multipotency. PMSCs were cultured in nondifferentiation or osteogenic differentiation conditions containing 15% FBS in room air (20% O 2 ) or low oxygen levels (1% O 2 ) for 14 days as in . For low/room treatment, PMSCs were cultured for 7 days in low oxygen and followed by 7 days in room air (shown in the third panel). Alizarin red or alkaline phosphatase staining was used to detect calcium deposition and enzyme expression changes as shown morphologically in (a) and quantified in (b) (two-way ANOVA, P < 0.05, N = 4); lowercase letter (a, b, and c) indicates significance between oxygen tension effects within nondifferentiation condition, uppercase letter (A, B, C) indicates within differentiation conditions. From immunoblots shown in Figure S2, protein levels were quantified in (c) OCT4, (d) SOX2, and (e) RUNX2 levels. Quantification levels shown were normalized to β -actin, a protein loading control (two-way ANOVA, P < 0.05, N = 3). ∗ indicates significance between room air and low oxygen tension; lowercase letter (a, b, and c) indicates significance between oxygen tension effects within nondifferentiation condition, uppercase letter (A, B, and C) indicates within differentiation conditions.

Journal: Stem Cells International

Article Title: Regulation of Osteogenic Differentiation of Placental-Derived Mesenchymal Stem Cells by Insulin-Like Growth Factors and Low Oxygen Tension

doi: 10.1155/2017/4576327

Figure Lengend Snippet: Effect of low oxygen tension preconditioning on PMSC osteogenic differentiation and multipotency. PMSCs were cultured in nondifferentiation or osteogenic differentiation conditions containing 15% FBS in room air (20% O 2 ) or low oxygen levels (1% O 2 ) for 14 days as in . For low/room treatment, PMSCs were cultured for 7 days in low oxygen and followed by 7 days in room air (shown in the third panel). Alizarin red or alkaline phosphatase staining was used to detect calcium deposition and enzyme expression changes as shown morphologically in (a) and quantified in (b) (two-way ANOVA, P < 0.05, N = 4); lowercase letter (a, b, and c) indicates significance between oxygen tension effects within nondifferentiation condition, uppercase letter (A, B, C) indicates within differentiation conditions. From immunoblots shown in Figure S2, protein levels were quantified in (c) OCT4, (d) SOX2, and (e) RUNX2 levels. Quantification levels shown were normalized to β -actin, a protein loading control (two-way ANOVA, P < 0.05, N = 3). ∗ indicates significance between room air and low oxygen tension; lowercase letter (a, b, and c) indicates significance between oxygen tension effects within nondifferentiation condition, uppercase letter (A, B, and C) indicates within differentiation conditions.

Techniques: Cell Culture, Staining, Expressing, Western Blot

PMSC multipotency and differentiation are regulated under low oxygen tension by IGFs. PMSCs were cultured for 14 days in osteogenic differentiation conditions containing 2% FBS in the presence or absence of 100 ng/mL of IGF-1 or IGF-2 in room air (20% O 2 ) or low oxygen levels (1% O 2 ). Treatments were stopped after 3, 7, and 14 days. Immunoblots, shown in Figure S3, were used to quantify the changes in protein levels of (a) OCT4, (b) SOX2, (c) RUNX2, and (d) OPN induced by IGFs over time. Quantification levels were normalized to β -actin, a protein loading control (two-way ANOVA, P < 0.05, N = 3). X indicates significance between different days without IGFs; # indicates significance of IGF addition compared with no IGFs in the same day.

Journal: Stem Cells International

Article Title: Regulation of Osteogenic Differentiation of Placental-Derived Mesenchymal Stem Cells by Insulin-Like Growth Factors and Low Oxygen Tension

doi: 10.1155/2017/4576327

Figure Lengend Snippet: PMSC multipotency and differentiation are regulated under low oxygen tension by IGFs. PMSCs were cultured for 14 days in osteogenic differentiation conditions containing 2% FBS in the presence or absence of 100 ng/mL of IGF-1 or IGF-2 in room air (20% O 2 ) or low oxygen levels (1% O 2 ). Treatments were stopped after 3, 7, and 14 days. Immunoblots, shown in Figure S3, were used to quantify the changes in protein levels of (a) OCT4, (b) SOX2, (c) RUNX2, and (d) OPN induced by IGFs over time. Quantification levels were normalized to β -actin, a protein loading control (two-way ANOVA, P < 0.05, N = 3). X indicates significance between different days without IGFs; # indicates significance of IGF addition compared with no IGFs in the same day.

Techniques: Cell Culture, Western Blot

Effect of oxygen tension and IGF-1 or IGF-2 on signaling downstream kinases of cell surface receptors during osteogenic differentiation. Similar to Figures and , PMSCs were cultured for 14 days in osteogenic differentiation conditions containing 2% FBS in the presence or absence of 100 ng/mL of IGF-1 or IGF-2 in room air (20% O 2 ) or low oxygen levels (1% O 2 ). Treatments were stopped after 3, 7, and 14 days. Immunoblots, in Figure S3, were used to quantify protein levels of (a) p-ERK1/2, (b) p-AKT, and (c) p-RUNX2 over the three days. These kinases/phosphoproteins were normalized to their total kinase/phosphoprotein level and β -actin (two-way ANOVA, P < 0.05, N = 3). X indicates significance between different days without IGFs; # indicates significance of IGF addition compared with no IGFs in the same day; ∗ indicates significance between room air and low oxygen tension.

Journal: Stem Cells International

Article Title: Regulation of Osteogenic Differentiation of Placental-Derived Mesenchymal Stem Cells by Insulin-Like Growth Factors and Low Oxygen Tension

doi: 10.1155/2017/4576327

Figure Lengend Snippet: Effect of oxygen tension and IGF-1 or IGF-2 on signaling downstream kinases of cell surface receptors during osteogenic differentiation. Similar to Figures and , PMSCs were cultured for 14 days in osteogenic differentiation conditions containing 2% FBS in the presence or absence of 100 ng/mL of IGF-1 or IGF-2 in room air (20% O 2 ) or low oxygen levels (1% O 2 ). Treatments were stopped after 3, 7, and 14 days. Immunoblots, in Figure S3, were used to quantify protein levels of (a) p-ERK1/2, (b) p-AKT, and (c) p-RUNX2 over the three days. These kinases/phosphoproteins were normalized to their total kinase/phosphoprotein level and β -actin (two-way ANOVA, P < 0.05, N = 3). X indicates significance between different days without IGFs; # indicates significance of IGF addition compared with no IGFs in the same day; ∗ indicates significance between room air and low oxygen tension.

Techniques: Cell Culture, Western Blot

PMSC differentiation is mediated via MEK1/2 and PI3K signaling and their inhibition effect on ERK1/2, AKT, and RUNX2 phosphorylation under low oxygen tension. PMSCs were cultured for 14 days in osteogenic differentiation conditions containing 2% FBS in room air (20% O 2 ) or low oxygen levels (1% O 2 ). During the 14 days, cells were continuously exposed to (5 μ M) U0126 or (10 μ M) LY294002 in differentiation media. Treatments were stopped at 14 days and stained with alizarin red to confirm (a) PMSC differentiation morphology changes with the inhibitors and quantified in (b) (two-way ANOVA, P < 0.05, N = 4). Immunoblots, shown in Figure S7, were used to quantify protein levels of (c) OCT4, (d) SOX2, (e) RUNX2, (f) OPN, (g) p-ERK1/2, (h) p-AKT, and (i) p-RUNX2 induced by signaling inhibition. Quantification levels were normalized to β -actin, a protein loading control; additionally, each phosphoprotein was normalized to its total protein (two-way ANOVA, P < 0.05, N = 3). ∗ indicates significance between room air and low oxygen tension; # indicates significance between DMSO control and inhibitor.

Journal: Stem Cells International

Article Title: Regulation of Osteogenic Differentiation of Placental-Derived Mesenchymal Stem Cells by Insulin-Like Growth Factors and Low Oxygen Tension

doi: 10.1155/2017/4576327

Figure Lengend Snippet: PMSC differentiation is mediated via MEK1/2 and PI3K signaling and their inhibition effect on ERK1/2, AKT, and RUNX2 phosphorylation under low oxygen tension. PMSCs were cultured for 14 days in osteogenic differentiation conditions containing 2% FBS in room air (20% O 2 ) or low oxygen levels (1% O 2 ). During the 14 days, cells were continuously exposed to (5 μ M) U0126 or (10 μ M) LY294002 in differentiation media. Treatments were stopped at 14 days and stained with alizarin red to confirm (a) PMSC differentiation morphology changes with the inhibitors and quantified in (b) (two-way ANOVA, P < 0.05, N = 4). Immunoblots, shown in Figure S7, were used to quantify protein levels of (c) OCT4, (d) SOX2, (e) RUNX2, (f) OPN, (g) p-ERK1/2, (h) p-AKT, and (i) p-RUNX2 induced by signaling inhibition. Quantification levels were normalized to β -actin, a protein loading control; additionally, each phosphoprotein was normalized to its total protein (two-way ANOVA, P < 0.05, N = 3). ∗ indicates significance between room air and low oxygen tension; # indicates significance between DMSO control and inhibitor.

Techniques: Inhibition, Cell Culture, Staining, Western Blot

PMSC osteogenic differentiation in the presence of IGFs and low oxygen tension. Summary diagram showing stem cell differentiation model in room air versus low oxygen. PMSCs can differentiate into osteogenic lineage only in room air, while maintaining PMSCs in low oxygen tension impedes this differentiation process. (A) In low oxygen tension, nondifferentiation conditions maintain self-renewal (higher proliferation) and IGF-2 can maintain higher expression of OCT4 and SOX2. (B) In differentiation media, PMSCs show an impeded differentiation with higher OCT4 and SOX2 and lowered RUNX2, which recovers following room air exposure. (C) In room air, spontaneous differentiation occurs with lower OCT4 and SOX2 and lower RUNX2. (D) In differentiation media, PMSCs fully differentiate towards osteoblastic progenitors with higher RUNX2 and OPN, which enhanced by IGF-1. Cell morphology and alizarin red staining increase as PMSCs lose multipotency and commit to the osteogenic lineage.

Journal: Stem Cells International

Article Title: Regulation of Osteogenic Differentiation of Placental-Derived Mesenchymal Stem Cells by Insulin-Like Growth Factors and Low Oxygen Tension

doi: 10.1155/2017/4576327

Figure Lengend Snippet: PMSC osteogenic differentiation in the presence of IGFs and low oxygen tension. Summary diagram showing stem cell differentiation model in room air versus low oxygen. PMSCs can differentiate into osteogenic lineage only in room air, while maintaining PMSCs in low oxygen tension impedes this differentiation process. (A) In low oxygen tension, nondifferentiation conditions maintain self-renewal (higher proliferation) and IGF-2 can maintain higher expression of OCT4 and SOX2. (B) In differentiation media, PMSCs show an impeded differentiation with higher OCT4 and SOX2 and lowered RUNX2, which recovers following room air exposure. (C) In room air, spontaneous differentiation occurs with lower OCT4 and SOX2 and lower RUNX2. (D) In differentiation media, PMSCs fully differentiate towards osteoblastic progenitors with higher RUNX2 and OPN, which enhanced by IGF-1. Cell morphology and alizarin red staining increase as PMSCs lose multipotency and commit to the osteogenic lineage.

Techniques: Cell Differentiation, Expressing, Staining

a , b Immunofluorescence shows overlapping of Tomato + and the expression of Runx2, Osterix, collagen X, MMP13, alkaline phosphatase, cleaved caspase 3 and LepR in POC (E18.5) and SOC (P15) of Lepr-cre; tdTomato mice. Arrowheads indicate the overlapping of Tomato + and the expression of indicated markers. HZ = Hypertrophic zone; POC = Primary ossification center; SOC = Secondary ossification center; ColX=collagen X; ALP = alkaline phosphatase; cleaved Casp 3=cleaved caspase 3. Scale bar, 100 μm.

Journal: Communications Biology

Article Title: PP2A in LepR+ mesenchymal stem cells contributes to embryonic and postnatal endochondral ossification through Runx2 dephosphorylation

doi: 10.1038/s42003-021-02175-1

Figure Lengend Snippet: a , b Immunofluorescence shows overlapping of Tomato + and the expression of Runx2, Osterix, collagen X, MMP13, alkaline phosphatase, cleaved caspase 3 and LepR in POC (E18.5) and SOC (P15) of Lepr-cre; tdTomato mice. Arrowheads indicate the overlapping of Tomato + and the expression of indicated markers. HZ = Hypertrophic zone; POC = Primary ossification center; SOC = Secondary ossification center; ColX=collagen X; ALP = alkaline phosphatase; cleaved Casp 3=cleaved caspase 3. Scale bar, 100 μm.

Article Snippet: The primary antibodies including anti-Ppp2r1a (Genetex GTX-102206, 1:500), anti-phospho Tyr307 PP2AC (Santa Cruz sc-271903, 1:100), anti-Runx2 (Cell Signaling 8486, 1:1000), anti-human phospho Ser451 Runx2 (Bioss bs-5685, 1:300), anti-PPARγ (ABclonal A0270, 1:500), anti-phospho Ser492 BRD4 (Millipore ABE1451, 1:500), anti-Osterix (Bioss bs-1110, 1:300), anti-collagen X (Abcam ab58632, 1:100), anti-MMP13 (Genetex GTX-100665, 1:500) and anti-GAPDH (Genetex GTX-100118, 1:5000) were used.

Techniques: Immunofluorescence, Expressing

Sections of E18.5 distal femur from Lepr-cre; Ppp2r1a fl/fl mice were subjected to immunostaining. a – d Immunohistochemistry (IHC) and immunofluorescence reveal that LepR + MSCs express Ppp2r1a and unphosphorylated (Y307) PP2AC in POC. a LepR IHC. b Ppp2r1a IHC. c Double immunofluorescence of LepR and Ppp2r1a. d Unphosphorylated (Y307) PP2AC IHC. e Illustration of conditional knockout of Ppp2r1a in LepR + MSCs. IHC reveals successful deletion of Ppp2r1a in LepR + MSCs in POC. IHC reveals that deletion of Ppp2r1a in LepR + MSCs leads to decreased expression of Ki67 ( f ) and hypertrophic markers, such as Runx2, Osterix, collagen X, MMP13 and alkaline phosphatase ( g ), and increased expression of Perilipin ( h ) in POC at E18.5. The rectangle box shows magnified picture. HZ = Hypertrophic zone; POC = Primary ossification center; ColX =collagen X; ALP = alkaline phosphatase. Scale bar, 100 μm.

Journal: Communications Biology

Article Title: PP2A in LepR+ mesenchymal stem cells contributes to embryonic and postnatal endochondral ossification through Runx2 dephosphorylation

doi: 10.1038/s42003-021-02175-1

Figure Lengend Snippet: Sections of E18.5 distal femur from Lepr-cre; Ppp2r1a fl/fl mice were subjected to immunostaining. a – d Immunohistochemistry (IHC) and immunofluorescence reveal that LepR + MSCs express Ppp2r1a and unphosphorylated (Y307) PP2AC in POC. a LepR IHC. b Ppp2r1a IHC. c Double immunofluorescence of LepR and Ppp2r1a. d Unphosphorylated (Y307) PP2AC IHC. e Illustration of conditional knockout of Ppp2r1a in LepR + MSCs. IHC reveals successful deletion of Ppp2r1a in LepR + MSCs in POC. IHC reveals that deletion of Ppp2r1a in LepR + MSCs leads to decreased expression of Ki67 ( f ) and hypertrophic markers, such as Runx2, Osterix, collagen X, MMP13 and alkaline phosphatase ( g ), and increased expression of Perilipin ( h ) in POC at E18.5. The rectangle box shows magnified picture. HZ = Hypertrophic zone; POC = Primary ossification center; ColX =collagen X; ALP = alkaline phosphatase. Scale bar, 100 μm.

Techniques: Immunostaining, Immunohistochemistry, Immunofluorescence, Knock-Out, Expressing

Sections of P15 distal femur from Lepr-cre; Ppp2r1a fl/fl mice were subjected to immunostaining. a–d Immunohistochemistry (IHC) and immunofluorescence reveal LepR + MSCs express Ppp2r1a and unphosphorylated (Y307) PP2AC in SOC. (a) LepR IHC. b Ppp2r1a IHC. c Double immunofluorescence of LepR and Ppp2r1a. d Unphosphorylated (Y307) PP2AC IHC. e IHC reveals successful deletion of Ppp2r1a in LepR + MSCs in SOC. IHC reveals that deletion of Ppp2r1a in LepR + MSCs leads to decreased expression of Ki67 ( f ) and hypertrophic markers, such as Runx2, Osterix, collagen X, MMP13 and alkaline phosphatase ( g ), and increased expression of Perilipin ( h ) in SOC at P15. HZ = Hypertrophic zone; POC = Primary ossification center; ColX =collagen X; ALP = alkaline phosphatase. Scale bar, 100 μm.

Journal: Communications Biology

Article Title: PP2A in LepR+ mesenchymal stem cells contributes to embryonic and postnatal endochondral ossification through Runx2 dephosphorylation

doi: 10.1038/s42003-021-02175-1

Figure Lengend Snippet: Sections of P15 distal femur from Lepr-cre; Ppp2r1a fl/fl mice were subjected to immunostaining. a–d Immunohistochemistry (IHC) and immunofluorescence reveal LepR + MSCs express Ppp2r1a and unphosphorylated (Y307) PP2AC in SOC. (a) LepR IHC. b Ppp2r1a IHC. c Double immunofluorescence of LepR and Ppp2r1a. d Unphosphorylated (Y307) PP2AC IHC. e IHC reveals successful deletion of Ppp2r1a in LepR + MSCs in SOC. IHC reveals that deletion of Ppp2r1a in LepR + MSCs leads to decreased expression of Ki67 ( f ) and hypertrophic markers, such as Runx2, Osterix, collagen X, MMP13 and alkaline phosphatase ( g ), and increased expression of Perilipin ( h ) in SOC at P15. HZ = Hypertrophic zone; POC = Primary ossification center; ColX =collagen X; ALP = alkaline phosphatase. Scale bar, 100 μm.

Techniques: Immunostaining, Immunohistochemistry, Immunofluorescence, Expressing

MSCs isolated from WT (WT MSCs) and Lepr-cre; Ppp2r1a fl/fl mice (PP2A KO MSCs) were subjected to analysis. a WT and PP2A KO MSCs exhibit no difference in morphology. Scale bar, 50 μm. b PP2A KO MSCs exhibit decreased proliferation. n = 3 independent experiments. c Represented flow cytometry plots of Annexin V-FITC/PI assay for apoptosis analysis of WT and PP2A KO MSCs. After FSC/SSC gating (left), MSCs are categorized into necrosis, late apoptosis, early apoptosis and live stages (right). The corresponding percentage are shown in bar chart. d WT and PP2A KO MSCs were induced for osteogenic differentiation and stained with Alizarin Red S at different time points. Scale bar, 1000 μm. e WT and PP2A KO MSCs were induced for adipogenic differentiation and stained with Oil red O at different time points. f WT and PP2A KO MSCs in micromass were induced for chondrogenic differentiation and stained with Alcian blue & nuclear fast red at different time points. Scale bar in e , f , 100 μm. g Quantification percentage of Alizarin red S staining, Oil red O staining and Alicante blue staining according to different differentiations respectively at each time points. n = 3 independent experiments. h qPCR analysis of transcript levels for genes associated with osteogenesis ( Runx2 , Alp ), adipogenesis ( Pparg , Adipoq ) and chondrogenesis ( Sox9 , Col2a1 ) differentiation in MSCs induced by differentiation medium at each time points. Transcript levels were normalized based on β-actin amplification. n = 3 independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001 as determined with Student’s t -test. Data are mean ± s.d.

Journal: Communications Biology

Article Title: PP2A in LepR+ mesenchymal stem cells contributes to embryonic and postnatal endochondral ossification through Runx2 dephosphorylation

doi: 10.1038/s42003-021-02175-1

Figure Lengend Snippet: MSCs isolated from WT (WT MSCs) and Lepr-cre; Ppp2r1a fl/fl mice (PP2A KO MSCs) were subjected to analysis. a WT and PP2A KO MSCs exhibit no difference in morphology. Scale bar, 50 μm. b PP2A KO MSCs exhibit decreased proliferation. n = 3 independent experiments. c Represented flow cytometry plots of Annexin V-FITC/PI assay for apoptosis analysis of WT and PP2A KO MSCs. After FSC/SSC gating (left), MSCs are categorized into necrosis, late apoptosis, early apoptosis and live stages (right). The corresponding percentage are shown in bar chart. d WT and PP2A KO MSCs were induced for osteogenic differentiation and stained with Alizarin Red S at different time points. Scale bar, 1000 μm. e WT and PP2A KO MSCs were induced for adipogenic differentiation and stained with Oil red O at different time points. f WT and PP2A KO MSCs in micromass were induced for chondrogenic differentiation and stained with Alcian blue & nuclear fast red at different time points. Scale bar in e , f , 100 μm. g Quantification percentage of Alizarin red S staining, Oil red O staining and Alicante blue staining according to different differentiations respectively at each time points. n = 3 independent experiments. h qPCR analysis of transcript levels for genes associated with osteogenesis ( Runx2 , Alp ), adipogenesis ( Pparg , Adipoq ) and chondrogenesis ( Sox9 , Col2a1 ) differentiation in MSCs induced by differentiation medium at each time points. Transcript levels were normalized based on β-actin amplification. n = 3 independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001 as determined with Student’s t -test. Data are mean ± s.d.

Techniques: Isolation, Flow Cytometry, Staining, Amplification

MSCs isolated from WT (WT MSCs) and Lepr-cre; Ppp2r1a fl/fl mice (PP2A KO MSCs) were subjected to analysis. a Representative western blot analysis of WT MSCs after osteogenic induction. b Representative western blot analysis of WT MSCs after adipogenic induction. c Western blot analysis of WT and PP2A KO MSCs. d Co-Immunoprecipitation study of cell lysates from WT MSCs with indicated induction for osteogenesis. IP with PP2A-specific or phospho Ser 472 Runx2-specific antibody and precipitates were probed for phospho Ser472 Runx2 and PP2A. e and f IHC analysis of inactive form phospho Ser472 Runx2 expression in hypertrophic chondrocyte of POC (E18.5) and SOC (P15) from Lepr-cre; Ppp2r1a fl/fl mice and relative WT littermate mice. g Quantification of phospho Ser472 Runx2 percentage in hypertrophic chondrocyte of POC (E18.5) and SOC (P15) from Lepr-cre; Ppp2r1a fl/fl mice and relative WT littermate mice. *** p < 0.001 as determined with Student’s t -test. Data are mean ± s.d. h Scheme representing regulation of MSC differentiation by PP2A and its substrates protein. Scale bar, 50 μm.

Journal: Communications Biology

Article Title: PP2A in LepR+ mesenchymal stem cells contributes to embryonic and postnatal endochondral ossification through Runx2 dephosphorylation

doi: 10.1038/s42003-021-02175-1

Figure Lengend Snippet: MSCs isolated from WT (WT MSCs) and Lepr-cre; Ppp2r1a fl/fl mice (PP2A KO MSCs) were subjected to analysis. a Representative western blot analysis of WT MSCs after osteogenic induction. b Representative western blot analysis of WT MSCs after adipogenic induction. c Western blot analysis of WT and PP2A KO MSCs. d Co-Immunoprecipitation study of cell lysates from WT MSCs with indicated induction for osteogenesis. IP with PP2A-specific or phospho Ser 472 Runx2-specific antibody and precipitates were probed for phospho Ser472 Runx2 and PP2A. e and f IHC analysis of inactive form phospho Ser472 Runx2 expression in hypertrophic chondrocyte of POC (E18.5) and SOC (P15) from Lepr-cre; Ppp2r1a fl/fl mice and relative WT littermate mice. g Quantification of phospho Ser472 Runx2 percentage in hypertrophic chondrocyte of POC (E18.5) and SOC (P15) from Lepr-cre; Ppp2r1a fl/fl mice and relative WT littermate mice. *** p < 0.001 as determined with Student’s t -test. Data are mean ± s.d. h Scheme representing regulation of MSC differentiation by PP2A and its substrates protein. Scale bar, 50 μm.

Techniques: Isolation, Western Blot, Immunoprecipitation, Expressing

Fig. 4. A-L: The microscopic images of immunohistochemical staining of tissues for TGF-β (A-E), Runx2 (F-J), and VEGF (K-O). The micrographs of the bare PMMA cement group are presented in A, F, and K; Micrographs of the estrogen-supplemented PMMA cement group in B, G, and L; Micrographs of the Bone-chips impregnated PMMA cement group in C, H, and M; Micrographs of the hydroxyapatite cement group in D, I and N; Micrographs of CPC group in E, J and O. Immunopositive structures in images are stained with chromogen in red color (arrows) and counter-staining was done with Mayer’s hematoxylin. Scale bars showing 200x and 400x magnifications.

Journal: Injury

Article Title: Modified and alternative bone cements can improve the induced membrane: Critical size bone defect model in rat femur.

doi: 10.1016/j.injury.2024.111627

Figure Lengend Snippet: Fig. 4. A-L: The microscopic images of immunohistochemical staining of tissues for TGF-β (A-E), Runx2 (F-J), and VEGF (K-O). The micrographs of the bare PMMA cement group are presented in A, F, and K; Micrographs of the estrogen-supplemented PMMA cement group in B, G, and L; Micrographs of the Bone-chips impregnated PMMA cement group in C, H, and M; Micrographs of the hydroxyapatite cement group in D, I and N; Micrographs of CPC group in E, J and O. Immunopositive structures in images are stained with chromogen in red color (arrows) and counter-staining was done with Mayer’s hematoxylin. Scale bars showing 200x and 400x magnifications.

Article Snippet: The Streptavidin-Biotin-Peroxidase method was performed by using the labeled monoclonal or polyclonal antibodies against TGF-β (sc-146, Santa Cruz Biotechnology), Runx2 (orb 102256; Biorbyt), and VEGF proteins (C-1; sc-7269).

Techniques: Immunohistochemical staining, Staining

Journal: Materials

Article Title: Bioactive Glasses in Periodontal Regeneration: Existing Strategies and Future Prospects—A Literature Review

doi: 10.3390/ma15062194

Figure Lengend Snippet: Summary of in vitro studies.

Article Snippet: Granel et al. [ ] , BG and PCL , Rat primary osteoblastic (RPO) cells Cell viability assay (XTT test) Cell proliferation (CyQUANT NF assay) SEM morphological evaluation Cell signaling (immunoassay for Runx2, FAK, phospho-FAK (Y397), GAPDH) Alkaline phosphatase activity assay.

Techniques: In Vitro, Activity Assay, Proliferation Assay, Gene Expression, Real-time Polymerase Chain Reaction, Expressing, BIA-KA, Mineralization Assay, Flow Cytometry, Cell Cycle Assay, MTT Assay, BrdU Staining, Staining, Western Blot, Reverse Transcription, Polymerase Chain Reaction, Alamar Blue Assay, Clinical Proteomics, CtB Assay, Adsorption, Viability Assay, MTS Assay, Modification, Dissolution, Migration, Microscopy, Membrane, XTT Assay, CyQUANT Assay, ALP Activity Assay, Cell Attachment Assay

Influence of obesity on RUNX2 in the presence and absence of EMD. ( a ) RUNX2 protein in normal-weight control animals, HFSD-fed animals, and HFSD-fed and EMD-treated animals. Representative immunohistochemistry images are shown. ( b ) Mean intensity of RUNX2 in normal-weight control animals, HFSD-fed animals, and HFSD-fed and EMD-treated animals. Bars show mean ± SEM; n = 5 animals/group; * significant ( p < 0.05) difference between groups. ( c ) Frequency distribution of different intensity categories for RUNX2 in normal-weight control animals, HFSD-fed animals, and HFSD-fed and EMD-treated animals. The intensity of immunohistochemical staining was assigned to five intensity categories (1 = very low, 2 = low, 3 = moderate, 4 = high, 5 = very high). EMD (enamel matrix derivative), HFSD (high fat, high sucrose diet), P (pulp), D (dentin), PDL (periodontal ligament), B (bone).

Journal: International Journal of Molecular Sciences

Article Title: Effects of Obesity on Bone Healing in Rats

doi: 10.3390/ijms222413339

Figure Lengend Snippet: Influence of obesity on RUNX2 in the presence and absence of EMD. ( a ) RUNX2 protein in normal-weight control animals, HFSD-fed animals, and HFSD-fed and EMD-treated animals. Representative immunohistochemistry images are shown. ( b ) Mean intensity of RUNX2 in normal-weight control animals, HFSD-fed animals, and HFSD-fed and EMD-treated animals. Bars show mean ± SEM; n = 5 animals/group; * significant ( p < 0.05) difference between groups. ( c ) Frequency distribution of different intensity categories for RUNX2 in normal-weight control animals, HFSD-fed animals, and HFSD-fed and EMD-treated animals. The intensity of immunohistochemical staining was assigned to five intensity categories (1 = very low, 2 = low, 3 = moderate, 4 = high, 5 = very high). EMD (enamel matrix derivative), HFSD (high fat, high sucrose diet), P (pulp), D (dentin), PDL (periodontal ligament), B (bone).

Article Snippet: Sections for RUNX2 detection were pretreated with pepsin at 37 °C for 20 min, followed by pre-blocking with 1× tris-buffered saline (Merck)/4% bovine serum albumin (Merck) at room temperature for 1 h. In the next step, the sections were incubated with a rabbit polyclonal antibody against RUNX2 (ab23981, abcam, Cambridge, UK) and a rabbit polyclonal anti-osteopontin antibody (ab8448, abcam).

Techniques: Control, Immunohistochemistry, Immunohistochemical staining, Staining

Colocalisation of Alizarin Red staining with osteogenic, inflammatory and apoptotic markers. Sequential sections of a valve leaflet stimulated media alone (left column) or 100 ng/mL LPS and 3 mM phosphate (right column) for 14-days. Histochemical staining for Alizarin Red (A,B) co-localizes with regions of immunohistochemical staining for osteocalcin (C,D) , RUNX2 (E,F) , NF-kB (G,H) and caspase 3 (I,J) . Scale bar, 500 μM.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Organ Culture Model of Aortic Valve Calcification

doi: 10.3389/fcvm.2021.734692

Figure Lengend Snippet: Colocalisation of Alizarin Red staining with osteogenic, inflammatory and apoptotic markers. Sequential sections of a valve leaflet stimulated media alone (left column) or 100 ng/mL LPS and 3 mM phosphate (right column) for 14-days. Histochemical staining for Alizarin Red (A,B) co-localizes with regions of immunohistochemical staining for osteocalcin (C,D) , RUNX2 (E,F) , NF-kB (G,H) and caspase 3 (I,J) . Scale bar, 500 μM.

Article Snippet: Slides were then incubated overnight in a moist chamber with antibodies against rabbit polyclonal RUNX2 at 1:200 (Abcam), mouse monoclonal Osteocalcin 1:600 (Abcam), rabbit polyclonal Osteopontin 1:500 (Chemicon), mouse monoclonal NF-kB 1:800 (BD transduction), Rabbit monoclonal cleaved caspase 3 1:200 (R&D systems).

Techniques: Staining, Immunohistochemical staining

Quantification of osteogenic, inflammatory and apoptotic markers in Alizarin Red positive and negative areas of valve leaflets. Percentage area staining of 1.1 mm 2 regions from non-calcified and calcified regions for (A) osteocalcin (* P = 0.012, T -Test; n = 4), (B) RUNX2 (** P < 0.001, T -Test; n = 4), (C) NF-kB (** P < 0.001, T -Test; n = 4) and (D) caspase 3 (** P < 0.001, T -Test; n = 4).

Journal: Frontiers in Cardiovascular Medicine

Article Title: Organ Culture Model of Aortic Valve Calcification

doi: 10.3389/fcvm.2021.734692

Figure Lengend Snippet: Quantification of osteogenic, inflammatory and apoptotic markers in Alizarin Red positive and negative areas of valve leaflets. Percentage area staining of 1.1 mm 2 regions from non-calcified and calcified regions for (A) osteocalcin (* P = 0.012, T -Test; n = 4), (B) RUNX2 (** P < 0.001, T -Test; n = 4), (C) NF-kB (** P < 0.001, T -Test; n = 4) and (D) caspase 3 (** P < 0.001, T -Test; n = 4).

Techniques: Staining

Effect of adenosine on the expression of osteogenic markers. Immunohistochemical staining in media alone (left column), 100 ng/mL LPS 3 μM phosphate treated (center column) and 100 ng/mL LPS 3 mM phosphate and 10 −5 M adenosine treated (right column) valve leaflets. Section were stained with RUNX2 (A–C) , osteopontin (D–F) and osteocalcin (G–I) . Scale bar, 100 μM.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Organ Culture Model of Aortic Valve Calcification

doi: 10.3389/fcvm.2021.734692

Figure Lengend Snippet: Effect of adenosine on the expression of osteogenic markers. Immunohistochemical staining in media alone (left column), 100 ng/mL LPS 3 μM phosphate treated (center column) and 100 ng/mL LPS 3 mM phosphate and 10 −5 M adenosine treated (right column) valve leaflets. Section were stained with RUNX2 (A–C) , osteopontin (D–F) and osteocalcin (G–I) . Scale bar, 100 μM.

Techniques: Expressing, Immunohistochemical staining, Staining

Inhibitory effect of adenosine on calcification and expression of osteogenic markers. Effect of increasing concentrations of adenosine (10 −8 -10 −5 M) on the expression of Alizarin Red in sections of valve leaflet response to 100 ng/mL and 3 mM phosphate (* P = 0.008, T -Test; n = 8 and ** P = 0.011, T -Test; n = 6) and representative images of sections stained with Alizarin Red imaged under polarized light for each treatment group (A) . Quantification of the area of positive staining in control (media alone), 100 ng/mL and 3 mM phosphate and 100 ng/mL & 3 mM phosphate with 10 −5 M adenosine for (B) osteocalcin (** P = 0.001, * P = 0.029, ANOVA; n = 11–15 areas from 3 valves), (C) osteopontin (** P < 0.001, * P = 0.029, ANOVA; n = 13–15 areas from 3 valves) and (D) RUNX2 (* P = 0.003, ** P = 0.001, ANOVA; n = 9–14 areas from 3 valves). (E) The mean area per measurement was similar in all 3 groups.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Organ Culture Model of Aortic Valve Calcification

doi: 10.3389/fcvm.2021.734692

Figure Lengend Snippet: Inhibitory effect of adenosine on calcification and expression of osteogenic markers. Effect of increasing concentrations of adenosine (10 −8 -10 −5 M) on the expression of Alizarin Red in sections of valve leaflet response to 100 ng/mL and 3 mM phosphate (* P = 0.008, T -Test; n = 8 and ** P = 0.011, T -Test; n = 6) and representative images of sections stained with Alizarin Red imaged under polarized light for each treatment group (A) . Quantification of the area of positive staining in control (media alone), 100 ng/mL and 3 mM phosphate and 100 ng/mL & 3 mM phosphate with 10 −5 M adenosine for (B) osteocalcin (** P = 0.001, * P = 0.029, ANOVA; n = 11–15 areas from 3 valves), (C) osteopontin (** P < 0.001, * P = 0.029, ANOVA; n = 13–15 areas from 3 valves) and (D) RUNX2 (* P = 0.003, ** P = 0.001, ANOVA; n = 9–14 areas from 3 valves). (E) The mean area per measurement was similar in all 3 groups.

Techniques: Expressing, Staining, Control

Figure 5. FO upregulates GSK-3β phosphorylation at Ser9 and thereby promotes the nuclear translocation of RUNX2, leading to calcium deposition. (A) MC3T3-E1 cells were treated with the indicated

Journal: Marine drugs

Article Title: Fermented Oyster Extract Promotes Insulin-Like Growth Factor-1-Mediated Osteogenesis and Growth Rate.

doi: 10.3390/md18090472

Figure Lengend Snippet: Figure 5. FO upregulates GSK-3β phosphorylation at Ser9 and thereby promotes the nuclear translocation of RUNX2, leading to calcium deposition. (A) MC3T3-E1 cells were treated with the indicated

Article Snippet: Antibodies against GSK-3β (sc-81462), phospho-GSK-3β at Ser9 (sc-37800), RUNX2 (sc-101145) and peroxidase-labeled anti-mouse immunoglobulins (sc-516102) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Phospho-proteomics, Translocation Assay

Figure 6. Transient knockdown of IGF-1Rαβ reduces calcium deposition concomitantly with a decrease in phosphorylated GSK-3β at Ser9. MC3T3-E1 cells were seeded at a density of 2000 cells/cm2 and transfected with silencing RNA for IGF-1Rαβ (siIGF-1Rαβ) 48 h before stimulation with 100 µg/mL FO or 2 mM GP. (A) Total mRNA was extracted, and reverse transcription polymerase chain reaction was performed. GAPDH was used as an internal control. (B) After 7 days, the cells were stained with alizarin red for calcium deposition, and images were captured. (C) Three days after treatment with FO, total proteins were isolated and Western blotting was performed to measure the protein level of phosphorylated GSK-3β at Ser9 (left). Total GSK-3β and β-actin were used as internal controls. The expression of IGF-1Rαβ and phosphorylated GSK-3β at Ser9 relative to GAPDH and total GSK-3β levels is illustrated (right). Signiﬁcant diﬀerences among the groups were determined using one-way ANOVA followed by Bonferroni correction. All data are presented as mean ± SEM (* p < 0.05 and *** p < 0.001 vs. untreated MC3T3-E1 cells). FO, fermented oyster (C. gigas) extract.

Journal: Marine drugs

Article Title: Fermented Oyster Extract Promotes Insulin-Like Growth Factor-1-Mediated Osteogenesis and Growth Rate.

doi: 10.3390/md18090472

Figure Lengend Snippet: Figure 6. Transient knockdown of IGF-1Rαβ reduces calcium deposition concomitantly with a decrease in phosphorylated GSK-3β at Ser9. MC3T3-E1 cells were seeded at a density of 2000 cells/cm2 and transfected with silencing RNA for IGF-1Rαβ (siIGF-1Rαβ) 48 h before stimulation with 100 µg/mL FO or 2 mM GP. (A) Total mRNA was extracted, and reverse transcription polymerase chain reaction was performed. GAPDH was used as an internal control. (B) After 7 days, the cells were stained with alizarin red for calcium deposition, and images were captured. (C) Three days after treatment with FO, total proteins were isolated and Western blotting was performed to measure the protein level of phosphorylated GSK-3β at Ser9 (left). Total GSK-3β and β-actin were used as internal controls. The expression of IGF-1Rαβ and phosphorylated GSK-3β at Ser9 relative to GAPDH and total GSK-3β levels is illustrated (right). Signiﬁcant diﬀerences among the groups were determined using one-way ANOVA followed by Bonferroni correction. All data are presented as mean ± SEM (* p < 0.05 and *** p < 0.001 vs. untreated MC3T3-E1 cells). FO, fermented oyster (C. gigas) extract.

Techniques: Knockdown, Transfection, Reverse Transcription, Polymerase Chain Reaction, Control, Staining, Isolation, Western Blot, Expressing

Figure 7. FO promotes bone formation and growth by activating the IGF-1/IGF-1R signaling pathway. FO upregulates growth-promoting genes such as insulin-like growth factor 1 (IGF-1), thereby promoting the release of IGF in osteoblast cells. The secreted IGF-1 binds to its speciﬁc receptor, IGF-1R, in an autocrine or paracrine manner and subsequently triggers the downstream signaling pathway. Once the IGF-1/IGF-1R complex is formed, it initiates the GSK-3β phosphorylation at Ser9. Phosphorylated GSK-3β subsequently releases RUNX2, which in turn promotes bone formation and growth. FO, fermented oyster (C. gigas) extract; IGF-1R, insulin-like growth factor-1 receptor; GSK-3β, glycogen synthase kinase-3β; RUNX2, runt-related transcription factor 2.

Journal: Marine drugs

Article Title: Fermented Oyster Extract Promotes Insulin-Like Growth Factor-1-Mediated Osteogenesis and Growth Rate.

doi: 10.3390/md18090472

Figure Lengend Snippet: Figure 7. FO promotes bone formation and growth by activating the IGF-1/IGF-1R signaling pathway. FO upregulates growth-promoting genes such as insulin-like growth factor 1 (IGF-1), thereby promoting the release of IGF in osteoblast cells. The secreted IGF-1 binds to its speciﬁc receptor, IGF-1R, in an autocrine or paracrine manner and subsequently triggers the downstream signaling pathway. Once the IGF-1/IGF-1R complex is formed, it initiates the GSK-3β phosphorylation at Ser9. Phosphorylated GSK-3β subsequently releases RUNX2, which in turn promotes bone formation and growth. FO, fermented oyster (C. gigas) extract; IGF-1R, insulin-like growth factor-1 receptor; GSK-3β, glycogen synthase kinase-3β; RUNX2, runt-related transcription factor 2.

Techniques: Phospho-proteomics

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