Fig. 1

Kindlin-2 loss in osteoblastic cells severely impairs skeletal response to intermittent PTH by affecting osteoblast and osteoclast function. a Three-dimensional (3D) images of micro-computerized tomography (μCT) of distal femurs from 3-month-old control (Cre-negative Kindlin-2^f/f) and Dmp1-Cre; Kindlin-2^f/f (cKO) female mice with and without PTH treatment for 28 d starting at the age of 3 months. b, c Quantitative analyses of the bone mineral density (BMD) and bone volume/tissue volume (BV/TV). N = 6 per group for both control and cKO. Results are expressed as mean ± standard deviation (s.d.). *P < 0.05, **P < 0.01, ***P < 0.001 (a–m). d–g Calcein double labeling. Images of the mineralized surface of the non-demineralized distal femoral sections (d). Scale bars: 20 μm. Quantitative analyses of measurement of the mineral apposition rate (MAR) (e), mineralizing surface per bone surface (MS/BS) (f), and bone formation rate (BFR) (g). N = 6 per group for both control and cKO. h–j Osteoclast formation in bone. Tibial sections of a were subjected to tartrate-resistant acid phosphatase (TRAP) staining. Scale bars: 50 μm. Quantitative analyses of the osteoclast surface/bone surface (Oc.S/BS) (i) and osteoclast number/bone perimeter (Oc.N/BPm) (j). N = 6 per group for both control and cKO. k–m Real-time RT-PCR (qPCR) analysis. Total RNA isolated from above control cKO bones was used for qPCR analysis for expression of Rankl and Opg mRNA, which was normalized to Gapdh mRNA. Experiments were independently repeated three times. n Immunofluorescence (IF) staining. Sections of tibial sections were subjected to IF staining with an antibody against osterix (Osx). Scale bars: 50 μm. Arrowheads indicate Osx-expressing osteoblasts