SETD2 loss of function in BMSCs showed decreased bone mass.

(A) 3D μ-CT images of trabecular bone of distal femurs isolated from 5-week-old male WT and Prx1-Cre, Setd2fl/fl mice. (B–F) μ-CT analysis of distal femur from 5 weeks old male WT and Prx1-Cre, Setd2fl/fl mice for bone volume per tissue volume, trabecular number, trabecular thickness, BMD, and trabecular separation. Results are presented as the mean ± SD. (n ≥ 6 per group, significant difference by Student t test; *p p Prx1-Cre, Setd2fl/fl mice. (H) μ-CT analysis of cortical thickness (Ct.Th) of middle shaft of femur from 5-week-old male WT and Prx1-Cre, Setd2fl/fl mice. (I) Immunofluorescence assay of Opn and Col1α1 of distal femur of 5-week-old WT and Prx1-Cre, Setd2fl/fl mice. Scale bar = 100μm. (J) Quantification of the area of Opn and Col1α1 staining. The y-axis was normalized with the area of WT mice. (K) Representative images of Von kossa staining of 5-week-old WT and Prx1-Cre, Setd2fl/fl mice. (L) Representative images of dual Calcein–Alizarin red S labeling of proximal tibia from WT and Prx1-Cre, Setd2fl/fl mice. Scale bar = 100 μm. (M) Quantification of MAR. Results are presented as the mean ± SD, n = 3 per group. Data used in the generation of this figure can be found in S1 Data. BMD, bone mineral density; BMSC, bone marrow mesenchymal stem cell; BV/TV, bone volume per tissue volume; Col1α1, Collagen type 1 alpha 1; MAR, mineral apposition rate; Opn, osteopontin; SETD2, SET-domain-containing 2; Tb. N, trabecular number; Tb. Sp, trabecular space; Tb. Th, trabecular thickness; WT, wild-type.