Fig. 2: Impaired mitochondrial calcium uptake, due to aging or Mcu loss, suppresses osteogenic differentiation.
From: Mcu regulates bone formation via mitochondrial calcium uptake and lineage allocation
a Schematic overview of mitochondrial calcium uptake measurement from mitochondrial isolation from pObs derived from young (8-week) or aged (60-week) mice. b Mitochondrial calcium uptake measured using Calcium Green-5N in isolated mitochondria treated with 5 μM calcium. Representative traces and quantification shown. c Immunoblot analysis of mitochondrial proteins in equal amounts of mitochondria isolated from each group. d ALP staining in MC3T3-E1 cells during osteogenic differentiation. e qPCR analysis of mitochondrial and osteogenic gene expression across different time points. f Immunoblot of OXPHOS subunits and osteogenic markers during MC3T3-E1 differentiation. g siRNA-mediated knockdown efficiency of Mcu confirmed by immunoblot. h Mitochondrial calcium uptake in isolated mitochondria from siRNA-treated cells. i Representative ALP staining on day 7 post-knockdown. j Von Kossa and Alizarin Red S staining after 14 days of differentiation. k Immunoblot analysis of osteogenic markers on day 7 post-knockdown. Data are presented as mean ± s.e.m. Statistical significance: *P < 0.05, ***P < 0.001, ****P < 0.0001.
