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
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