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mouse osteocyte cell line mlo-y4 cells ( )  (Absolute Biotech Inc)

 
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    Absolute Biotech Inc mouse osteocyte cell line mlo-y4 cells ( )
    Effect of PTH and Pi on <t>MLO-Y4</t> cell death. (A–C) Cell death levels in MLO-Y4 cells cultured with (A) Pi or the mixture of Pi and PTH (Pi + PTH); (B) PTH, and (C) FGF23 for 1 to 3 days (n = 4). Cell death levels in cells treated with Pi were much higher than those in controls in a dose and time-dependent manner. Cell death levels in cells treated with Pi + PTH were higher than those in the cells treated with Pi. Conversely, cell death levels in cells treated with PTH or FGF23 were similar to those in the control cells. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with control). (D) Localization of Pit1 and Pit2 (green) in MLO-Y4 cells after 6 and 12 hours of Pi treatment (n = 3; nuclei stained with DAPI). Pit1 and Pit2 immunoreactivities were localized around the cell membrane in the MLO-Y4 cells without Pi treatment. In Pi-treated MLO-Y4 cells, Pit1 and Pit2 immunoreactivities were observed in the granular structure inside the cells. (E-J) Expression levels of Pit1 and Pit2 in MLO-Y4 cells treated with indicated compounds for 3, 6, 12, and 24 hours (n = 3). Pit1 expression levels in MLO-Y4 cells increased significantly after 3 hours in all Pi, PTH, and FGF23 treatments; thereafter, the expression levels decreased with increased duration of treatment. Pit1 expression levels were elevated more substantially by Pi or PTH treatment than by FGF23 treatment. Pit2 expression in MLO-Y4 cells slightly increased when the cells were treated with Pi and PTH. The Pit2 expression did not markedly change by FGF23 treatment. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with the 3-hour controls). ###, P < 0.001; ##, P < 0.01; #, P < 0.05 (compared with the 6-hour controls). †††, P < 0.001; ††, P < 0.01; †, P < 0.05 (compared with the 12-hour controls). §§§, P < 0.001; §§, P < 0.01; §, P < 0.05 (compared with the 24-hour controls). Data are shown as the mean ± SE; differences with P < 0.05 were considered significant. Abbreviations: FGF23, fibroblast growth factor 23; Pi, inorganic phosphate.
    Mouse Osteocyte Cell Line Mlo Y4 Cells ( ), supplied by Absolute Biotech Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+osteocyte+cell+line+mlo-y4+cells+%28+%29/pmc09939342-91-0-13?v=Absolute+Biotech+Inc
    Average 90 stars, based on 1 article reviews
    mouse osteocyte cell line mlo-y4 cells ( ) - by Bioz Stars, 2026-08
    90/100 stars

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    1) Product Images from "Evocalcet Rescues Secondary Hyperparathyroidism-driven Cortical Porosity in CKD Male Rats"

    Article Title: Evocalcet Rescues Secondary Hyperparathyroidism-driven Cortical Porosity in CKD Male Rats

    Journal: Endocrinology

    doi: 10.1210/endocr/bqad022

    Effect of PTH and Pi on MLO-Y4 cell death. (A–C) Cell death levels in MLO-Y4 cells cultured with (A) Pi or the mixture of Pi and PTH (Pi + PTH); (B) PTH, and (C) FGF23 for 1 to 3 days (n = 4). Cell death levels in cells treated with Pi were much higher than those in controls in a dose and time-dependent manner. Cell death levels in cells treated with Pi + PTH were higher than those in the cells treated with Pi. Conversely, cell death levels in cells treated with PTH or FGF23 were similar to those in the control cells. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with control). (D) Localization of Pit1 and Pit2 (green) in MLO-Y4 cells after 6 and 12 hours of Pi treatment (n = 3; nuclei stained with DAPI). Pit1 and Pit2 immunoreactivities were localized around the cell membrane in the MLO-Y4 cells without Pi treatment. In Pi-treated MLO-Y4 cells, Pit1 and Pit2 immunoreactivities were observed in the granular structure inside the cells. (E-J) Expression levels of Pit1 and Pit2 in MLO-Y4 cells treated with indicated compounds for 3, 6, 12, and 24 hours (n = 3). Pit1 expression levels in MLO-Y4 cells increased significantly after 3 hours in all Pi, PTH, and FGF23 treatments; thereafter, the expression levels decreased with increased duration of treatment. Pit1 expression levels were elevated more substantially by Pi or PTH treatment than by FGF23 treatment. Pit2 expression in MLO-Y4 cells slightly increased when the cells were treated with Pi and PTH. The Pit2 expression did not markedly change by FGF23 treatment. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with the 3-hour controls). ###, P < 0.001; ##, P < 0.01; #, P < 0.05 (compared with the 6-hour controls). †††, P < 0.001; ††, P < 0.01; †, P < 0.05 (compared with the 12-hour controls). §§§, P < 0.001; §§, P < 0.01; §, P < 0.05 (compared with the 24-hour controls). Data are shown as the mean ± SE; differences with P < 0.05 were considered significant. Abbreviations: FGF23, fibroblast growth factor 23; Pi, inorganic phosphate.
    Figure Legend Snippet: Effect of PTH and Pi on MLO-Y4 cell death. (A–C) Cell death levels in MLO-Y4 cells cultured with (A) Pi or the mixture of Pi and PTH (Pi + PTH); (B) PTH, and (C) FGF23 for 1 to 3 days (n = 4). Cell death levels in cells treated with Pi were much higher than those in controls in a dose and time-dependent manner. Cell death levels in cells treated with Pi + PTH were higher than those in the cells treated with Pi. Conversely, cell death levels in cells treated with PTH or FGF23 were similar to those in the control cells. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with control). (D) Localization of Pit1 and Pit2 (green) in MLO-Y4 cells after 6 and 12 hours of Pi treatment (n = 3; nuclei stained with DAPI). Pit1 and Pit2 immunoreactivities were localized around the cell membrane in the MLO-Y4 cells without Pi treatment. In Pi-treated MLO-Y4 cells, Pit1 and Pit2 immunoreactivities were observed in the granular structure inside the cells. (E-J) Expression levels of Pit1 and Pit2 in MLO-Y4 cells treated with indicated compounds for 3, 6, 12, and 24 hours (n = 3). Pit1 expression levels in MLO-Y4 cells increased significantly after 3 hours in all Pi, PTH, and FGF23 treatments; thereafter, the expression levels decreased with increased duration of treatment. Pit1 expression levels were elevated more substantially by Pi or PTH treatment than by FGF23 treatment. Pit2 expression in MLO-Y4 cells slightly increased when the cells were treated with Pi and PTH. The Pit2 expression did not markedly change by FGF23 treatment. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with the 3-hour controls). ###, P < 0.001; ##, P < 0.01; #, P < 0.05 (compared with the 6-hour controls). †††, P < 0.001; ††, P < 0.01; †, P < 0.05 (compared with the 12-hour controls). §§§, P < 0.001; §§, P < 0.01; §, P < 0.05 (compared with the 24-hour controls). Data are shown as the mean ± SE; differences with P < 0.05 were considered significant. Abbreviations: FGF23, fibroblast growth factor 23; Pi, inorganic phosphate.

    Techniques Used: Cell Culture, Staining, Expressing



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    mouse osteocyte cell line mlo-y4 cells ( )  (Absolute Biotech Inc)
    90
    Absolute Biotech Inc mouse osteocyte cell line mlo-y4 cells ( )
    Effect of PTH and Pi on <t>MLO-Y4</t> cell death. (A–C) Cell death levels in MLO-Y4 cells cultured with (A) Pi or the mixture of Pi and PTH (Pi + PTH); (B) PTH, and (C) FGF23 for 1 to 3 days (n = 4). Cell death levels in cells treated with Pi were much higher than those in controls in a dose and time-dependent manner. Cell death levels in cells treated with Pi + PTH were higher than those in the cells treated with Pi. Conversely, cell death levels in cells treated with PTH or FGF23 were similar to those in the control cells. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with control). (D) Localization of Pit1 and Pit2 (green) in MLO-Y4 cells after 6 and 12 hours of Pi treatment (n = 3; nuclei stained with DAPI). Pit1 and Pit2 immunoreactivities were localized around the cell membrane in the MLO-Y4 cells without Pi treatment. In Pi-treated MLO-Y4 cells, Pit1 and Pit2 immunoreactivities were observed in the granular structure inside the cells. (E-J) Expression levels of Pit1 and Pit2 in MLO-Y4 cells treated with indicated compounds for 3, 6, 12, and 24 hours (n = 3). Pit1 expression levels in MLO-Y4 cells increased significantly after 3 hours in all Pi, PTH, and FGF23 treatments; thereafter, the expression levels decreased with increased duration of treatment. Pit1 expression levels were elevated more substantially by Pi or PTH treatment than by FGF23 treatment. Pit2 expression in MLO-Y4 cells slightly increased when the cells were treated with Pi and PTH. The Pit2 expression did not markedly change by FGF23 treatment. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with the 3-hour controls). ###, P < 0.001; ##, P < 0.01; #, P < 0.05 (compared with the 6-hour controls). †††, P < 0.001; ††, P < 0.01; †, P < 0.05 (compared with the 12-hour controls). §§§, P < 0.001; §§, P < 0.01; §, P < 0.05 (compared with the 24-hour controls). Data are shown as the mean ± SE; differences with P < 0.05 were considered significant. Abbreviations: FGF23, fibroblast growth factor 23; Pi, inorganic phosphate.
    Mouse Osteocyte Cell Line Mlo Y4 Cells ( ), supplied by Absolute Biotech Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+osteocyte+cell+line+mlo-y4+cells+%28+%29/pmc09939342-91-0-13?v=Absolute+Biotech+Inc
    Average 90 stars, based on 1 article reviews
    mouse osteocyte cell line mlo-y4 cells ( ) - by Bioz Stars, 2026-08
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    mlo y4 cells mouse osteocyte cell line mlo y4 cells  (Absolute Biotech Inc)
    86
    Absolute Biotech Inc mlo y4 cells mouse osteocyte cell line mlo y4 cells
    Effect of PTH and Pi on <t>MLO-Y4</t> cell death. (A–C) Cell death levels in MLO-Y4 cells cultured with (A) Pi or the mixture of Pi and PTH (Pi + PTH); (B) PTH, and (C) FGF23 for 1 to 3 days (n = 4). Cell death levels in cells treated with Pi were much higher than those in controls in a dose and time-dependent manner. Cell death levels in cells treated with Pi + PTH were higher than those in the cells treated with Pi. Conversely, cell death levels in cells treated with PTH or FGF23 were similar to those in the control cells. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with control). (D) Localization of Pit1 and Pit2 (green) in MLO-Y4 cells after 6 and 12 hours of Pi treatment (n = 3; nuclei stained with DAPI). Pit1 and Pit2 immunoreactivities were localized around the cell membrane in the MLO-Y4 cells without Pi treatment. In Pi-treated MLO-Y4 cells, Pit1 and Pit2 immunoreactivities were observed in the granular structure inside the cells. (E-J) Expression levels of Pit1 and Pit2 in MLO-Y4 cells treated with indicated compounds for 3, 6, 12, and 24 hours (n = 3). Pit1 expression levels in MLO-Y4 cells increased significantly after 3 hours in all Pi, PTH, and FGF23 treatments; thereafter, the expression levels decreased with increased duration of treatment. Pit1 expression levels were elevated more substantially by Pi or PTH treatment than by FGF23 treatment. Pit2 expression in MLO-Y4 cells slightly increased when the cells were treated with Pi and PTH. The Pit2 expression did not markedly change by FGF23 treatment. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with the 3-hour controls). ###, P < 0.001; ##, P < 0.01; #, P < 0.05 (compared with the 6-hour controls). †††, P < 0.001; ††, P < 0.01; †, P < 0.05 (compared with the 12-hour controls). §§§, P < 0.001; §§, P < 0.01; §, P < 0.05 (compared with the 24-hour controls). Data are shown as the mean ± SE; differences with P < 0.05 were considered significant. Abbreviations: FGF23, fibroblast growth factor 23; Pi, inorganic phosphate.
    Mlo Y4 Cells Mouse Osteocyte Cell Line Mlo Y4 Cells, supplied by Absolute Biotech 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/mouse+osteocyte+cell+line+mlo-y4+cells+%28+%29/pm36718587-101-10-23?v=Absolute+Biotech+Inc
    Average 86 stars, based on 1 article reviews
    mlo y4 cells mouse osteocyte cell line mlo y4 cells - by Bioz Stars, 2026-08
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    mouse osteocyte cell line mlo-y4  (Procell Inc)
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    Procell Inc mouse osteocyte cell line mlo-y4
    Effect of PTH and Pi on <t>MLO-Y4</t> cell death. (A–C) Cell death levels in MLO-Y4 cells cultured with (A) Pi or the mixture of Pi and PTH (Pi + PTH); (B) PTH, and (C) FGF23 for 1 to 3 days (n = 4). Cell death levels in cells treated with Pi were much higher than those in controls in a dose and time-dependent manner. Cell death levels in cells treated with Pi + PTH were higher than those in the cells treated with Pi. Conversely, cell death levels in cells treated with PTH or FGF23 were similar to those in the control cells. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with control). (D) Localization of Pit1 and Pit2 (green) in MLO-Y4 cells after 6 and 12 hours of Pi treatment (n = 3; nuclei stained with DAPI). Pit1 and Pit2 immunoreactivities were localized around the cell membrane in the MLO-Y4 cells without Pi treatment. In Pi-treated MLO-Y4 cells, Pit1 and Pit2 immunoreactivities were observed in the granular structure inside the cells. (E-J) Expression levels of Pit1 and Pit2 in MLO-Y4 cells treated with indicated compounds for 3, 6, 12, and 24 hours (n = 3). Pit1 expression levels in MLO-Y4 cells increased significantly after 3 hours in all Pi, PTH, and FGF23 treatments; thereafter, the expression levels decreased with increased duration of treatment. Pit1 expression levels were elevated more substantially by Pi or PTH treatment than by FGF23 treatment. Pit2 expression in MLO-Y4 cells slightly increased when the cells were treated with Pi and PTH. The Pit2 expression did not markedly change by FGF23 treatment. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with the 3-hour controls). ###, P < 0.001; ##, P < 0.01; #, P < 0.05 (compared with the 6-hour controls). †††, P < 0.001; ††, P < 0.01; †, P < 0.05 (compared with the 12-hour controls). §§§, P < 0.001; §§, P < 0.01; §, P < 0.05 (compared with the 24-hour controls). Data are shown as the mean ± SE; differences with P < 0.05 were considered significant. Abbreviations: FGF23, fibroblast growth factor 23; Pi, inorganic phosphate.
    Mouse Osteocyte Cell Line Mlo Y4, supplied by Procell Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+osteocyte+cell+line+mlo-y4+cells+%28+%29/pm33933560-129-0-8?v=Procell+Inc
    Average 90 stars, based on 1 article reviews
    mouse osteocyte cell line mlo-y4 - by Bioz Stars, 2026-08
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    immortalized mouse primary osteocyte-like cell line mlo-y4  (Asahi Kasei Corporation)
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    Asahi Kasei Corporation immortalized mouse primary osteocyte-like cell line mlo-y4
    Effect of PTH and Pi on <t>MLO-Y4</t> cell death. (A–C) Cell death levels in MLO-Y4 cells cultured with (A) Pi or the mixture of Pi and PTH (Pi + PTH); (B) PTH, and (C) FGF23 for 1 to 3 days (n = 4). Cell death levels in cells treated with Pi were much higher than those in controls in a dose and time-dependent manner. Cell death levels in cells treated with Pi + PTH were higher than those in the cells treated with Pi. Conversely, cell death levels in cells treated with PTH or FGF23 were similar to those in the control cells. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with control). (D) Localization of Pit1 and Pit2 (green) in MLO-Y4 cells after 6 and 12 hours of Pi treatment (n = 3; nuclei stained with DAPI). Pit1 and Pit2 immunoreactivities were localized around the cell membrane in the MLO-Y4 cells without Pi treatment. In Pi-treated MLO-Y4 cells, Pit1 and Pit2 immunoreactivities were observed in the granular structure inside the cells. (E-J) Expression levels of Pit1 and Pit2 in MLO-Y4 cells treated with indicated compounds for 3, 6, 12, and 24 hours (n = 3). Pit1 expression levels in MLO-Y4 cells increased significantly after 3 hours in all Pi, PTH, and FGF23 treatments; thereafter, the expression levels decreased with increased duration of treatment. Pit1 expression levels were elevated more substantially by Pi or PTH treatment than by FGF23 treatment. Pit2 expression in MLO-Y4 cells slightly increased when the cells were treated with Pi and PTH. The Pit2 expression did not markedly change by FGF23 treatment. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with the 3-hour controls). ###, P < 0.001; ##, P < 0.01; #, P < 0.05 (compared with the 6-hour controls). †††, P < 0.001; ††, P < 0.01; †, P < 0.05 (compared with the 12-hour controls). §§§, P < 0.001; §§, P < 0.01; §, P < 0.05 (compared with the 24-hour controls). Data are shown as the mean ± SE; differences with P < 0.05 were considered significant. Abbreviations: FGF23, fibroblast growth factor 23; Pi, inorganic phosphate.
    Immortalized Mouse Primary Osteocyte Like Cell Line Mlo Y4, supplied by Asahi Kasei Corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+osteocyte+cell+line+mlo-y4+cells+%28+%29/10__1299_slash_jbse__6__101-35-2-26?v=Asahi+Kasei+Corporation
    Average 90 stars, based on 1 article reviews
    immortalized mouse primary osteocyte-like cell line mlo-y4 - by Bioz Stars, 2026-08
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    mouse primary osteocyte-like cell line mlo-y4  (Asahi Kasei Corporation)
    90
    Asahi Kasei Corporation mouse primary osteocyte-like cell line mlo-y4
    Effect of PTH and Pi on <t>MLO-Y4</t> cell death. (A–C) Cell death levels in MLO-Y4 cells cultured with (A) Pi or the mixture of Pi and PTH (Pi + PTH); (B) PTH, and (C) FGF23 for 1 to 3 days (n = 4). Cell death levels in cells treated with Pi were much higher than those in controls in a dose and time-dependent manner. Cell death levels in cells treated with Pi + PTH were higher than those in the cells treated with Pi. Conversely, cell death levels in cells treated with PTH or FGF23 were similar to those in the control cells. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with control). (D) Localization of Pit1 and Pit2 (green) in MLO-Y4 cells after 6 and 12 hours of Pi treatment (n = 3; nuclei stained with DAPI). Pit1 and Pit2 immunoreactivities were localized around the cell membrane in the MLO-Y4 cells without Pi treatment. In Pi-treated MLO-Y4 cells, Pit1 and Pit2 immunoreactivities were observed in the granular structure inside the cells. (E-J) Expression levels of Pit1 and Pit2 in MLO-Y4 cells treated with indicated compounds for 3, 6, 12, and 24 hours (n = 3). Pit1 expression levels in MLO-Y4 cells increased significantly after 3 hours in all Pi, PTH, and FGF23 treatments; thereafter, the expression levels decreased with increased duration of treatment. Pit1 expression levels were elevated more substantially by Pi or PTH treatment than by FGF23 treatment. Pit2 expression in MLO-Y4 cells slightly increased when the cells were treated with Pi and PTH. The Pit2 expression did not markedly change by FGF23 treatment. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with the 3-hour controls). ###, P < 0.001; ##, P < 0.01; #, P < 0.05 (compared with the 6-hour controls). †††, P < 0.001; ††, P < 0.01; †, P < 0.05 (compared with the 12-hour controls). §§§, P < 0.001; §§, P < 0.01; §, P < 0.05 (compared with the 24-hour controls). Data are shown as the mean ± SE; differences with P < 0.05 were considered significant. Abbreviations: FGF23, fibroblast growth factor 23; Pi, inorganic phosphate.
    Mouse Primary Osteocyte Like Cell Line Mlo Y4, supplied by Asahi Kasei Corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+osteocyte+cell+line+mlo-y4+cells+%28+%29/10__1299_slash_jbse__3__13-52-1-25?v=Asahi+Kasei+Corporation
    Average 90 stars, based on 1 article reviews
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    Effect of PTH and Pi on MLO-Y4 cell death. (A–C) Cell death levels in MLO-Y4 cells cultured with (A) Pi or the mixture of Pi and PTH (Pi + PTH); (B) PTH, and (C) FGF23 for 1 to 3 days (n = 4). Cell death levels in cells treated with Pi were much higher than those in controls in a dose and time-dependent manner. Cell death levels in cells treated with Pi + PTH were higher than those in the cells treated with Pi. Conversely, cell death levels in cells treated with PTH or FGF23 were similar to those in the control cells. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with control). (D) Localization of Pit1 and Pit2 (green) in MLO-Y4 cells after 6 and 12 hours of Pi treatment (n = 3; nuclei stained with DAPI). Pit1 and Pit2 immunoreactivities were localized around the cell membrane in the MLO-Y4 cells without Pi treatment. In Pi-treated MLO-Y4 cells, Pit1 and Pit2 immunoreactivities were observed in the granular structure inside the cells. (E-J) Expression levels of Pit1 and Pit2 in MLO-Y4 cells treated with indicated compounds for 3, 6, 12, and 24 hours (n = 3). Pit1 expression levels in MLO-Y4 cells increased significantly after 3 hours in all Pi, PTH, and FGF23 treatments; thereafter, the expression levels decreased with increased duration of treatment. Pit1 expression levels were elevated more substantially by Pi or PTH treatment than by FGF23 treatment. Pit2 expression in MLO-Y4 cells slightly increased when the cells were treated with Pi and PTH. The Pit2 expression did not markedly change by FGF23 treatment. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with the 3-hour controls). ###, P < 0.001; ##, P < 0.01; #, P < 0.05 (compared with the 6-hour controls). †††, P < 0.001; ††, P < 0.01; †, P < 0.05 (compared with the 12-hour controls). §§§, P < 0.001; §§, P < 0.01; §, P < 0.05 (compared with the 24-hour controls). Data are shown as the mean ± SE; differences with P < 0.05 were considered significant. Abbreviations: FGF23, fibroblast growth factor 23; Pi, inorganic phosphate.

    Journal: Endocrinology

    Article Title: Evocalcet Rescues Secondary Hyperparathyroidism-driven Cortical Porosity in CKD Male Rats

    doi: 10.1210/endocr/bqad022

    Figure Lengend Snippet: Effect of PTH and Pi on MLO-Y4 cell death. (A–C) Cell death levels in MLO-Y4 cells cultured with (A) Pi or the mixture of Pi and PTH (Pi + PTH); (B) PTH, and (C) FGF23 for 1 to 3 days (n = 4). Cell death levels in cells treated with Pi were much higher than those in controls in a dose and time-dependent manner. Cell death levels in cells treated with Pi + PTH were higher than those in the cells treated with Pi. Conversely, cell death levels in cells treated with PTH or FGF23 were similar to those in the control cells. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with control). (D) Localization of Pit1 and Pit2 (green) in MLO-Y4 cells after 6 and 12 hours of Pi treatment (n = 3; nuclei stained with DAPI). Pit1 and Pit2 immunoreactivities were localized around the cell membrane in the MLO-Y4 cells without Pi treatment. In Pi-treated MLO-Y4 cells, Pit1 and Pit2 immunoreactivities were observed in the granular structure inside the cells. (E-J) Expression levels of Pit1 and Pit2 in MLO-Y4 cells treated with indicated compounds for 3, 6, 12, and 24 hours (n = 3). Pit1 expression levels in MLO-Y4 cells increased significantly after 3 hours in all Pi, PTH, and FGF23 treatments; thereafter, the expression levels decreased with increased duration of treatment. Pit1 expression levels were elevated more substantially by Pi or PTH treatment than by FGF23 treatment. Pit2 expression in MLO-Y4 cells slightly increased when the cells were treated with Pi and PTH. The Pit2 expression did not markedly change by FGF23 treatment. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (compared with the 3-hour controls). ###, P < 0.001; ##, P < 0.01; #, P < 0.05 (compared with the 6-hour controls). †††, P < 0.001; ††, P < 0.01; †, P < 0.05 (compared with the 12-hour controls). §§§, P < 0.001; §§, P < 0.01; §, P < 0.05 (compared with the 24-hour controls). Data are shown as the mean ± SE; differences with P < 0.05 were considered significant. Abbreviations: FGF23, fibroblast growth factor 23; Pi, inorganic phosphate.

    Article Snippet: Mouse osteocyte cell line MLO-Y4 cells ( ) (2 × 10 5 cells; Kerafast, Inc., Boston, MA, USA) were seeded in a T25 flask containing an α-MEM (FUJIFILM Wako Pure Chemical Co.) containing 2.5% fetal bovine serum (Hyclone # SH30396; Cytive, Tokyo, Japan) and 2.5% calf serum (Hyclone # SH30072, Cytive) and then incubated in a 5% CO 2 incubator for 24 hours.

    Techniques: Cell Culture, Staining, Expressing