Fig. 2: Padi2 deficiency impaired osteoblast differentiation.

A Representative view of 4-week-old female Cont and Padi2-KO^Col1 mice. B Representative micro-CT images of the distal femur from 4-week-old female Cont and Padi2^Col1 mice in midsagittal and coronal views. Scale bar: 500 μm. C Histomorphometric analysis of 3D micro-CT of the distal femur from data 4-week-old female Cont (n = 10) and Padi2^Col1 (n = 6) mice. D IHC for PADI2 of distal femur from 4-week-old Cont and Padi2^Col1 mice. Three independent experiments with three biological replicates for each group. Scale bar: 200 μm, 100 μm. E Representative images of H&E staining for distal femur from 4-week-old Cont and Padi2^Col1 mice. Three independent experiments with three biological replicates for each group. Scale bar: 500 μm, 200 μm. F Relative mRNA expression of Padi2 and bone marker genes in Cont and KO primary calvaria osteoblasts (pOBs) cultured or not (Day0) in osteogenic medium for each indicated day as determined by RT-qPCR. Three independent experiments with three biological replicates for each group. G ALP and ARS staining were performed in Cont, Het, and KO primary calvaria OBs cultivated in an osteogenic medium for 5 days and 21 days, respectively. Representative images from three independent experiments with two biological replicates for each group. H TRAP staining of distal femurs from 4-week-old Cont and Padi2^Col1 mice. The boxed region is shown in higher magnification (bottom). Three independent experiments with three biological replicates for each group. Scale bar: 500 μm, 200 μm. Data are expressed as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. N.S, not significant. ND not determined.