Fig. 1: The expression of REPIN1 increases in osteoporosis patients, and iron overload impairs osteogenic function in vitro.

A, B The expression levels of REPIN1 in bone tissues from osteoporosis patients were analyzed by western blotting. C The mRNA levels of REPIN1 from osteoporosis patients were analyzed. D, E The expression level of REPIN1 in bone tissues of mice was analyzed by western blotting. F The mRNA levels of Repin1 were analyzed by qRT‒PCR. G Representative micro-CT reconstruction images of trabecular bone under the distal femur growth plate in the control and FAC groups. H, I The expression levels of REPIN1 in BMSCs exposed to FAC (0–200 μmol/L) were analyzed by western blotting. J The mRNA levels of Repin1 in BMSCs exposed to FAC (0–200 μmol/L) were analyzed by qRT‒PCR. K Perl’s Prussian blue staining was used to stain the iron particles in cells. The blue-stained iron particles are indicated by arrows. L Semiquantitative evaluation of iron particles after Perl’s Prussian blue staining. M ARS staining at 21 days. N Semiquantitative evaluation of the ARS recovery ratio (fold change). O ALP staining at 7 days. P Relative ALP expression level (fold change). (Cells in G–J were exposed to FAC at concentrations of 200 μmol/L for 48 h before examination. Values are shown as the means ± SDs **p < 0.01 and ***p < 0.005, n = 3 per group all these studies were performed at least three biological replicates).