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integrin β3  (Santa Cruz Biotechnology)


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    Structured Review

    Santa Cruz Biotechnology integrin β3
    AZD3965 inhibits key osteoclast marker protein expression in vitro. (A) Protein expression of <t>integrin-β3,</t> NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after 5-day treatment with RANKL and AZD3965 (0, 5, 10 μM). (B–F) Quantification of osteoclast marker protein expression with specified intervention (all normalized to β-actin). (G) NFATc1 expression and nuclear translocation detected via immunofluorescence staining (scale bar = 50 μm). (H) Quantification of NFATc1 mean fluorescence intensity in the nucleus. (I) Protein expression of integrin-β3, NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after treatment with RANKL and AZD3965 at different time points. (J–N) Quantification of osteoclast marker protein expression on the specified day (all normalized to β-actin). N = 3. ##P < 0.01, ###P < 0.001, ####P < 0.0001, compared to control group. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001, compared to the RANKL alone treatment group.
    Integrin β3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 336 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Designed bone-targeting ROS-responsive nanoplatform for precision glycolysis inhibition in postmenopausal osteoporosis"

    Article Title: Designed bone-targeting ROS-responsive nanoplatform for precision glycolysis inhibition in postmenopausal osteoporosis

    Journal: Bioactive Materials

    doi: 10.1016/j.bioactmat.2025.11.032

    AZD3965 inhibits key osteoclast marker protein expression in vitro. (A) Protein expression of integrin-β3, NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after 5-day treatment with RANKL and AZD3965 (0, 5, 10 μM). (B–F) Quantification of osteoclast marker protein expression with specified intervention (all normalized to β-actin). (G) NFATc1 expression and nuclear translocation detected via immunofluorescence staining (scale bar = 50 μm). (H) Quantification of NFATc1 mean fluorescence intensity in the nucleus. (I) Protein expression of integrin-β3, NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after treatment with RANKL and AZD3965 at different time points. (J–N) Quantification of osteoclast marker protein expression on the specified day (all normalized to β-actin). N = 3. ##P < 0.01, ###P < 0.001, ####P < 0.0001, compared to control group. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001, compared to the RANKL alone treatment group.
    Figure Legend Snippet: AZD3965 inhibits key osteoclast marker protein expression in vitro. (A) Protein expression of integrin-β3, NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after 5-day treatment with RANKL and AZD3965 (0, 5, 10 μM). (B–F) Quantification of osteoclast marker protein expression with specified intervention (all normalized to β-actin). (G) NFATc1 expression and nuclear translocation detected via immunofluorescence staining (scale bar = 50 μm). (H) Quantification of NFATc1 mean fluorescence intensity in the nucleus. (I) Protein expression of integrin-β3, NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after treatment with RANKL and AZD3965 at different time points. (J–N) Quantification of osteoclast marker protein expression on the specified day (all normalized to β-actin). N = 3. ##P < 0.01, ###P < 0.001, ####P < 0.0001, compared to control group. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001, compared to the RANKL alone treatment group.

    Techniques Used: Marker, Expressing, In Vitro, Western Blot, Translocation Assay, Immunofluorescence, Staining, Fluorescence, Control

    PH/DPA@A inhibits RANKL-induced osteoclast formation and bone resorption function in vitro. (A) Protein expression of integrin-β3, NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after 5-day treatment with RANKL and PH/DPA@A in five groups. (B–F) Quantification of osteoclast marker protein expression with indicated intervention (all normalized to β-actin). (G) TRAcP staining to assess osteoclast differentiation of BMMs in five groups. (H) Quantification of TRAcP staining in five groups. (I, J) F-actin staining and quantification of F-actin ring area in five groups. (K–L) Bone slice assay using mature osteoclasts and quantification of resorption area in five groups. Scale bar = 200 μm. N = 3. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001.
    Figure Legend Snippet: PH/DPA@A inhibits RANKL-induced osteoclast formation and bone resorption function in vitro. (A) Protein expression of integrin-β3, NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after 5-day treatment with RANKL and PH/DPA@A in five groups. (B–F) Quantification of osteoclast marker protein expression with indicated intervention (all normalized to β-actin). (G) TRAcP staining to assess osteoclast differentiation of BMMs in five groups. (H) Quantification of TRAcP staining in five groups. (I, J) F-actin staining and quantification of F-actin ring area in five groups. (K–L) Bone slice assay using mature osteoclasts and quantification of resorption area in five groups. Scale bar = 200 μm. N = 3. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001.

    Techniques Used: In Vitro, Expressing, Western Blot, Marker, Staining

    Related Articles

    Marker:

    Article Title: Designed bone-targeting ROS-responsive nanoplatform for precision glycolysis inhibition in postmenopausal osteoporosis
    Article Snippet: Cell culture supplies, including α-MEM and fetal bovine serum (FBS), were sourced from Thermo Fisher Scientific in Carlsbad, United States. .. Antibodies against osteoclast marker proteins, such as c-Fos, CTSK, integrin β3, and NFATc1, were procured from Santa Cruz (San Jose, United States). .. Moreover, antibodies targeting p-P38, P38, p-JNK, JNK, p-ERK, ERK, p-P65, P65, and IκB-α were sourced from Cell Signaling Technology, Danvers, United States.

    other:

    Article Title: Extracellular matrix protein signaling promotes multi-step cancer vasculogenic mimicry formation
    Article Snippet: 3-D Culture Matrix Rat Collagen I (3447-020-01) and 3-D Culture Matrix Laminin I (3446-005-01) were purchased from R&D Systems (Minneapolis, MN). siRNA against Integrin β1 (sc-35674), primary antibodies Integrin β1 (P5D2, sc-13590), Cortactin (H-5, sc-55579) and Integrin β3 (2C9.G2, sc-46655) were purchased from Santa Cruz Biotechnology (Dallas, TX).

    Article Title: Extracellular matrix protein signaling promotes multi-step cancer vasculogenic mimicry formation.
    Article Snippet: 3-D Culture Matrix Rat Collagen I (3447-020-01) and 3-D Culture Matrix Laminin I (3446-005-01) were purchased from R&D Systems (Minneapolis, MN). siRNA against Integrin β1 (sc-35674), primary antibodies Integrin β1 (P5D2, sc-13590), Cortactin (H-5, sc-55579) and Integrin β3 (2C9.G2, sc-46655) were purchased from Santa Cruz Biotechnology (Dallas, TX).

    Incubation:

    Article Title: Bone-on-a-chip recapitulates dynamic bone remodeling.
    Article Snippet: Bone remodeling is governed by the basic multicellular unit (BMU) and plays a crucial role in both normal bone function and pathological conditions (e.g. osteoporosis).. Unfortunately, our understanding of the bone remodeling process is limited as current models do not allow dynamic events in the BMU to be systematically and spatiotemporally analyzed.. Leveraging microfluidic and stem cell technologies, we developed an bone-on-a-chip model that enables the dynamic co-culture of osteoblasts (OBs) and osteoclasts (OCs) in a spatiotemporal context.

    Article Title: Integrin β1 Mediates Mechanosensitive Regulation of Human Trabecular Meshwork Cell Functions in Response to Substrate Stiffness
    Article Snippet: .. Cells were incubated overnight at 4 °C in a dark humid chamber with primary antibodies against integrin β1 (1:200, MAB2000; Sigma), integrin β3 (1:200, sc-365679; Santa Cruz Biotechnology, USA), integrin β5 (1:200, sc-398214; Santa Cruz Biotechnology), integrin αV (1:200, sc-9969; Santa Cruz Biotechnology), FN (1:200, F7387; Sigma), LN (1:200, sc59732; Santa Cruz Biotechnology) or VIM (1:200, sc-32322; Santa Cruz Biotechnology). .. After washing, cells were incubated with Alexa Fluor-conjugated secondary antibodies (1:200; ZF-0312; Zhongshan Golden Bridge, China) for 2 h at room temperature.

    Blocking Assay:

    Article Title: Bone-on-a-chip recapitulates dynamic bone remodeling.
    Article Snippet: Bone remodeling is governed by the basic multicellular unit (BMU) and plays a crucial role in both normal bone function and pathological conditions (e.g. osteoporosis).. Unfortunately, our understanding of the bone remodeling process is limited as current models do not allow dynamic events in the BMU to be systematically and spatiotemporally analyzed.. Leveraging microfluidic and stem cell technologies, we developed an bone-on-a-chip model that enables the dynamic co-culture of osteoblasts (OBs) and osteoclasts (OCs) in a spatiotemporal context.



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    AZD3965 inhibits key osteoclast marker protein expression in vitro. (A) Protein expression of <t>integrin-β3,</t> NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after 5-day treatment with RANKL and AZD3965 (0, 5, 10 μM). (B–F) Quantification of osteoclast marker protein expression with specified intervention (all normalized to β-actin). (G) NFATc1 expression and nuclear translocation detected via immunofluorescence staining (scale bar = 50 μm). (H) Quantification of NFATc1 mean fluorescence intensity in the nucleus. (I) Protein expression of integrin-β3, NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after treatment with RANKL and AZD3965 at different time points. (J–N) Quantification of osteoclast marker protein expression on the specified day (all normalized to β-actin). N = 3. ##P < 0.01, ###P < 0.001, ####P < 0.0001, compared to control group. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001, compared to the RANKL alone treatment group.
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    Image Search Results


    Characterization of human-derived hOsteo4 subclones. (A) Phase contrast images of 4 hOsteo4 subclones, that is, C7, D8, E9, and C10; (B) cell viability test by CCK8 assay for 4 hOsteo4 subclones and MLO-Y4 cells; N = 3 for each group; (C) immunofluorescence staining of DMP1, FGF23, and sclerostin of 4 hOsteo4 subclones, MLO-Y4 and MC3T3 cells; IgG group were MC3T3 cells and served as a negative background control; (D) western-blot results for Dmp1, Sclerostin, E11, integrin β1, integrin β3, ALP, and Col-I protein expressions from hOsteo4 subclones, MLO-Y4 cells and MC-3 T3 cells; tubulin served as a loading control.

    Journal: JBMR Plus

    Article Title: Establishment and characterization of a human pre-osteocyte cell line: hOsteo4-E9

    doi: 10.1093/jbmrpl/ziaf163

    Figure Lengend Snippet: Characterization of human-derived hOsteo4 subclones. (A) Phase contrast images of 4 hOsteo4 subclones, that is, C7, D8, E9, and C10; (B) cell viability test by CCK8 assay for 4 hOsteo4 subclones and MLO-Y4 cells; N = 3 for each group; (C) immunofluorescence staining of DMP1, FGF23, and sclerostin of 4 hOsteo4 subclones, MLO-Y4 and MC3T3 cells; IgG group were MC3T3 cells and served as a negative background control; (D) western-blot results for Dmp1, Sclerostin, E11, integrin β1, integrin β3, ALP, and Col-I protein expressions from hOsteo4 subclones, MLO-Y4 cells and MC-3 T3 cells; tubulin served as a loading control.

    Article Snippet: Integrin β3 , CST , 13166S , 1:1000 , .

    Techniques: Derivative Assay, CCK-8 Assay, Immunofluorescence, Staining, Control, Western Blot

    Subclone hOsteo4-E9 cells underwent dramatic morphological changes upon FSS treatment. (A) Phase contrast images showed the morphological changes of hOsteo4 subclones in static and FSS conditions; (B) quantitative analysis of percentages of dendritic cells of 4 hOsteo4 subclones with and without FSS; N = 3 for each group; (C) WB results of p-FAK, integrin β3, and kindlin-2 protein changes of hOsteo4-E9 subclone under different FSS stimuli; (D) phase contrast images of cell shape changes of hOsteo4-E9 subclone under different FSS stimuli; (E-G) statistical analysis of percentages of dendritic cells, number of dendrites per cell and the longest dendritic length per cell of hOsteo4-E9 subclone under FSS stimuli; quantitative results were analyzed from 3 independent experiments; (H) IF staining of F-actin cytoskeleton and DAPI nuclei in hOsteo4-E9 cells with and without (static) FSS stimuli; (I) IF images stained with F-actin, p-FAK, tubulin, and Cx43 in hOsteo4-E9 cells with and without (static) FSS stimuli. Arrowheads in figures indicated the direction of FSS forces. Results are expressed as mean ± SD. n. s. p > .05; * p < .05; ** p < .01; *** p < .001.

    Journal: JBMR Plus

    Article Title: Establishment and characterization of a human pre-osteocyte cell line: hOsteo4-E9

    doi: 10.1093/jbmrpl/ziaf163

    Figure Lengend Snippet: Subclone hOsteo4-E9 cells underwent dramatic morphological changes upon FSS treatment. (A) Phase contrast images showed the morphological changes of hOsteo4 subclones in static and FSS conditions; (B) quantitative analysis of percentages of dendritic cells of 4 hOsteo4 subclones with and without FSS; N = 3 for each group; (C) WB results of p-FAK, integrin β3, and kindlin-2 protein changes of hOsteo4-E9 subclone under different FSS stimuli; (D) phase contrast images of cell shape changes of hOsteo4-E9 subclone under different FSS stimuli; (E-G) statistical analysis of percentages of dendritic cells, number of dendrites per cell and the longest dendritic length per cell of hOsteo4-E9 subclone under FSS stimuli; quantitative results were analyzed from 3 independent experiments; (H) IF staining of F-actin cytoskeleton and DAPI nuclei in hOsteo4-E9 cells with and without (static) FSS stimuli; (I) IF images stained with F-actin, p-FAK, tubulin, and Cx43 in hOsteo4-E9 cells with and without (static) FSS stimuli. Arrowheads in figures indicated the direction of FSS forces. Results are expressed as mean ± SD. n. s. p > .05; * p < .05; ** p < .01; *** p < .001.

    Article Snippet: Integrin β3 , CST , 13166S , 1:1000 , .

    Techniques: Staining

    hOsteo4-E9 cells showed cytoskeleton remodeling with enhanced proteins expressions of mechanosensitive genes similar to MLO-Y4 cells in response to FSS treatment. (A) Phase contrast images of MLO-Y4 and hOsteo4-E9 cells under static/0, 1.5, and 3.5 dynes/cm 2 FSS; (B-E) quantitative analysis of dendritic percentages, number of dendrites per cell, the longest dendritic length in single cell and spreading area changes in MLO-Y4 and hOsteo4-E9 cells under different FSS stimuli; (F) SEM images of MLO-Y4 and hOsteo4-E9 cells with or without FSS; (G) F-actin staining of MLO-Y4 and hOsteo4-E9 cells with or without FSS; (H, I) quantitative analysis of the length of secondary dendrites and the number of secondary dendrites per cell in MLO-Y4 and hOsteo4-E9 cells with or without FSS; (J) western-blot results for p-FAK, kindlin-2, integrin β3, Cx43, Dmp1, and Sclerostin in MLO-Y4, and hOsteo4-E9 cells under different FSS stimuli; tubulin served as a loading control; (K-P) statistical analysis of protein expressions of p-FAK, kindlin-2, integrin β3, Cx43, Dmp1, and Sclerostin in MLO-Y4 and hOsteo4-E9 cells under different FSS stimuli. N = 3 for each group. Arrowheads in figures indicated the direction of FSS forces. Results are expressed as mean ± SD. n.s. p > .05; * p < .05; ** p < .01; *** p < .001.

    Journal: JBMR Plus

    Article Title: Establishment and characterization of a human pre-osteocyte cell line: hOsteo4-E9

    doi: 10.1093/jbmrpl/ziaf163

    Figure Lengend Snippet: hOsteo4-E9 cells showed cytoskeleton remodeling with enhanced proteins expressions of mechanosensitive genes similar to MLO-Y4 cells in response to FSS treatment. (A) Phase contrast images of MLO-Y4 and hOsteo4-E9 cells under static/0, 1.5, and 3.5 dynes/cm 2 FSS; (B-E) quantitative analysis of dendritic percentages, number of dendrites per cell, the longest dendritic length in single cell and spreading area changes in MLO-Y4 and hOsteo4-E9 cells under different FSS stimuli; (F) SEM images of MLO-Y4 and hOsteo4-E9 cells with or without FSS; (G) F-actin staining of MLO-Y4 and hOsteo4-E9 cells with or without FSS; (H, I) quantitative analysis of the length of secondary dendrites and the number of secondary dendrites per cell in MLO-Y4 and hOsteo4-E9 cells with or without FSS; (J) western-blot results for p-FAK, kindlin-2, integrin β3, Cx43, Dmp1, and Sclerostin in MLO-Y4, and hOsteo4-E9 cells under different FSS stimuli; tubulin served as a loading control; (K-P) statistical analysis of protein expressions of p-FAK, kindlin-2, integrin β3, Cx43, Dmp1, and Sclerostin in MLO-Y4 and hOsteo4-E9 cells under different FSS stimuli. N = 3 for each group. Arrowheads in figures indicated the direction of FSS forces. Results are expressed as mean ± SD. n.s. p > .05; * p < .05; ** p < .01; *** p < .001.

    Article Snippet: Integrin β3 , CST , 13166S , 1:1000 , .

    Techniques: Single Cell, Staining, Western Blot, Control

    RNA-sequencing data revealed that hOsteo4-E9 cells not only share some similarity with MLO-Y4 cells, but also have distinct transcription profiles in response to FSS. (A) PCA plot of gene expression variability of human hOsteo4-E9, murine MLO-Y4, and murine long bone-derived osteocytes; (B) Venn diagram of detected genes in hOsteo4-E9 and MLO-Y4 cells; (C) Venn diagram of detected genes in 4 experimental groups, that is, E9_F, E9_S, Y4_F, and Y4_S; (D-F) top 15 of KEGG enriched pathways in all detected genes, hOsteo4-E9 specific and MLO-Y4 specific genes; (G) clustering heat map highlighted the variations of FA-associated gene expression patterns between 4 experimental groups; (H-M) qPCR validation of gene expression of integrin αν, integrin α5, integrin α3, integrin β5, integrin β7, and EMP1 in 4 experimental groups. N = 3 for each group. Results are expressed as mean ± SD. n.s. p > .05; * p < .05; ** p < .01; *** p < .001.

    Journal: JBMR Plus

    Article Title: Establishment and characterization of a human pre-osteocyte cell line: hOsteo4-E9

    doi: 10.1093/jbmrpl/ziaf163

    Figure Lengend Snippet: RNA-sequencing data revealed that hOsteo4-E9 cells not only share some similarity with MLO-Y4 cells, but also have distinct transcription profiles in response to FSS. (A) PCA plot of gene expression variability of human hOsteo4-E9, murine MLO-Y4, and murine long bone-derived osteocytes; (B) Venn diagram of detected genes in hOsteo4-E9 and MLO-Y4 cells; (C) Venn diagram of detected genes in 4 experimental groups, that is, E9_F, E9_S, Y4_F, and Y4_S; (D-F) top 15 of KEGG enriched pathways in all detected genes, hOsteo4-E9 specific and MLO-Y4 specific genes; (G) clustering heat map highlighted the variations of FA-associated gene expression patterns between 4 experimental groups; (H-M) qPCR validation of gene expression of integrin αν, integrin α5, integrin α3, integrin β5, integrin β7, and EMP1 in 4 experimental groups. N = 3 for each group. Results are expressed as mean ± SD. n.s. p > .05; * p < .05; ** p < .01; *** p < .001.

    Article Snippet: Integrin β3 , CST , 13166S , 1:1000 , .

    Techniques: RNA Sequencing, Gene Expression, Derivative Assay, Biomarker Discovery

    AZD3965 inhibits key osteoclast marker protein expression in vitro. (A) Protein expression of integrin-β3, NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after 5-day treatment with RANKL and AZD3965 (0, 5, 10 μM). (B–F) Quantification of osteoclast marker protein expression with specified intervention (all normalized to β-actin). (G) NFATc1 expression and nuclear translocation detected via immunofluorescence staining (scale bar = 50 μm). (H) Quantification of NFATc1 mean fluorescence intensity in the nucleus. (I) Protein expression of integrin-β3, NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after treatment with RANKL and AZD3965 at different time points. (J–N) Quantification of osteoclast marker protein expression on the specified day (all normalized to β-actin). N = 3. ##P < 0.01, ###P < 0.001, ####P < 0.0001, compared to control group. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001, compared to the RANKL alone treatment group.

    Journal: Bioactive Materials

    Article Title: Designed bone-targeting ROS-responsive nanoplatform for precision glycolysis inhibition in postmenopausal osteoporosis

    doi: 10.1016/j.bioactmat.2025.11.032

    Figure Lengend Snippet: AZD3965 inhibits key osteoclast marker protein expression in vitro. (A) Protein expression of integrin-β3, NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after 5-day treatment with RANKL and AZD3965 (0, 5, 10 μM). (B–F) Quantification of osteoclast marker protein expression with specified intervention (all normalized to β-actin). (G) NFATc1 expression and nuclear translocation detected via immunofluorescence staining (scale bar = 50 μm). (H) Quantification of NFATc1 mean fluorescence intensity in the nucleus. (I) Protein expression of integrin-β3, NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after treatment with RANKL and AZD3965 at different time points. (J–N) Quantification of osteoclast marker protein expression on the specified day (all normalized to β-actin). N = 3. ##P < 0.01, ###P < 0.001, ####P < 0.0001, compared to control group. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001, compared to the RANKL alone treatment group.

    Article Snippet: Antibodies against osteoclast marker proteins, such as c-Fos, CTSK, integrin β3, and NFATc1, were procured from Santa Cruz (San Jose, United States).

    Techniques: Marker, Expressing, In Vitro, Western Blot, Translocation Assay, Immunofluorescence, Staining, Fluorescence, Control

    PH/DPA@A inhibits RANKL-induced osteoclast formation and bone resorption function in vitro. (A) Protein expression of integrin-β3, NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after 5-day treatment with RANKL and PH/DPA@A in five groups. (B–F) Quantification of osteoclast marker protein expression with indicated intervention (all normalized to β-actin). (G) TRAcP staining to assess osteoclast differentiation of BMMs in five groups. (H) Quantification of TRAcP staining in five groups. (I, J) F-actin staining and quantification of F-actin ring area in five groups. (K–L) Bone slice assay using mature osteoclasts and quantification of resorption area in five groups. Scale bar = 200 μm. N = 3. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001.

    Journal: Bioactive Materials

    Article Title: Designed bone-targeting ROS-responsive nanoplatform for precision glycolysis inhibition in postmenopausal osteoporosis

    doi: 10.1016/j.bioactmat.2025.11.032

    Figure Lengend Snippet: PH/DPA@A inhibits RANKL-induced osteoclast formation and bone resorption function in vitro. (A) Protein expression of integrin-β3, NFATc1, MMP9, CTSK and c-Fos detected via Western blotting after 5-day treatment with RANKL and PH/DPA@A in five groups. (B–F) Quantification of osteoclast marker protein expression with indicated intervention (all normalized to β-actin). (G) TRAcP staining to assess osteoclast differentiation of BMMs in five groups. (H) Quantification of TRAcP staining in five groups. (I, J) F-actin staining and quantification of F-actin ring area in five groups. (K–L) Bone slice assay using mature osteoclasts and quantification of resorption area in five groups. Scale bar = 200 μm. N = 3. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001.

    Article Snippet: Antibodies against osteoclast marker proteins, such as c-Fos, CTSK, integrin β3, and NFATc1, were procured from Santa Cruz (San Jose, United States).

    Techniques: In Vitro, Expressing, Western Blot, Marker, Staining

    Thrombogenicity evaluation of endothelialized scaffolds ( A ) schematic of the experimental design of ex vivo blood perfusion. ( B ) Gross appearance of DLS, endothelialized, and LXW7 endothelialized scaffolds after blood perfusion, showing markedly reduced clot deposition in the LXW7 group (yellow circles highlight thrombi). Scale bar = 2 cm. ( C ) IF staining of platelet marker integrin αIIb (red) with DAPI (blue) showing reduced platelet adhesion in endothelialized and LXW7endothelialized scaffolds compared to DLS. Scale bar = 100 µm. ( D ) Quantification of fluorescence intensity confirming significantly decreased platelet adhesion in LXW7 endothelialized scaffolds ( n = 4 fields, * p < 0.05 vs. DLS). ( E ) Time-dependent changes in platelet count (%) in the blood perfusate during ex vivo blood perfusion ( n = 3 samples each time point, * p < 0.05 vs. DLS).

    Journal: Journal of Functional Biomaterials

    Article Title: LXW7 Peptide Modification of Acellular Liver Scaffolds Improves Endothelialization and Hemocompatibility in Bioengineered Liver

    doi: 10.3390/jfb17030122

    Figure Lengend Snippet: Thrombogenicity evaluation of endothelialized scaffolds ( A ) schematic of the experimental design of ex vivo blood perfusion. ( B ) Gross appearance of DLS, endothelialized, and LXW7 endothelialized scaffolds after blood perfusion, showing markedly reduced clot deposition in the LXW7 group (yellow circles highlight thrombi). Scale bar = 2 cm. ( C ) IF staining of platelet marker integrin αIIb (red) with DAPI (blue) showing reduced platelet adhesion in endothelialized and LXW7endothelialized scaffolds compared to DLS. Scale bar = 100 µm. ( D ) Quantification of fluorescence intensity confirming significantly decreased platelet adhesion in LXW7 endothelialized scaffolds ( n = 4 fields, * p < 0.05 vs. DLS). ( E ) Time-dependent changes in platelet count (%) in the blood perfusate during ex vivo blood perfusion ( n = 3 samples each time point, * p < 0.05 vs. DLS).

    Article Snippet: For IF staining, sections were permeabilized with 0.1% Triton X-100 for 15 min and blocked with 2% bovine serum albumin (Sigma-Aldrich) for 45 min. Tissue sections were incubated overnight with primary antibodies including anti-human CD31 (1:100, MA5-13188, Invitrogen), Albumin (1:200, PA5-89332, Invitrogen), anti-Integrin αIIb (1:100, sc-21783, Santa Cruz Biotechnology, Dallas, TX, USA), and anti-TGF-β1 (1:200, ab170874, Abcam).

    Techniques: Ex Vivo, Staining, Marker, Fluorescence