Fig. 5
Becn1 of the megakaryocytic lineage cells negatively regulates bone mass in males. a, b Micro-CT analysis of Becn1f/f and Becn1f/f;Pf4-iCre mice. c Representative calcein double-labeling images with quantification of the mineral apposition rate (MAR). d Representative image and statistical analysis of HE staining of the femur. e Three-point mechanical testing of bone strength and stiffness. f Diagram of the whole bone marrow transplantation mouse model. g HE staining of the femurs of transplant recipient mice (Becn1+/+−Becn1+/+, n = 2; Becn1+/+−Becn1–/–, n = 4; Becn1–/–−Becn1+/+, n = 2; Becn1–/– −Becn1–/–, n = 3). h, i Micro-CT analysis of the transplanted mice. j Analysis of the Becn1 protein expression level in platelets by western blotting. β-Actin was used as a loading control. Left, representative western blot; right, statistical graph showing the relative expression levels of Becn1. k Percentages of GFP-positive platelets in rLV and rLV-Becn1 mice. l Statistical analysis of the percentages of GFP-positive MKPs, MKs, and platelets in rLV and rLV-Becn1 mice. m, n Micro-CT analysis of rLV and rLV-Becn1 mice. (rLV: Mice with megakaryocytic lineage-specific overexpression of the vector; rLV-Becn1: Mice with megakaryocytic lineage-specific overexpression of Becn1. Data are means ± SEMs. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.000 1
