Fig. 4

Mice lacking SHP2 in Bglap ⁺ osteoblastic cells display elevated local osteoclastogenesis and bone resorption. a Femoral frozen sections from the indicated 13-week-old mice show the activity of tartrate-resistant acid phosphatase (TRAP). Note the markedly increased TRAP activity in the cortical bone of SHP2 mutants compared to control mice. The quantitative data are provided in Fig. S2d. Scale bar: 100 μm. b Femoral frozen sections show the transcript abundance of the indicated osteoclastic genes in the metaphysis and diaphyseal cortical bone using RNAScope® technology (n = 3–4). Scale bar: 100 μm. c Femoral frozen sections show the transcript abundance of RANKL (Tnfsf11) and OPG (Tnfrsf11b) in the diaphyseal cortical bone using RNAScope® assays (n = 3). Scale bar: 100 μm. d Western blots show the expression of RANKL and SHP2 and the phosphorylation of STAT3 Y705 in osteoblast cell lines derived from the indicated mice. e Femoral frozen sections immunostained with antibodies against pSTAT3 Y705 demonstrate STAT3 activation in the OBs (arrows) (n = 4). Scale bar: 100 μm. f Western blots show the abundance and activation of STAT3 in SHP2-sufficient and SHP2-deficient OBs upon IL6 stimulation. g Western blots show the abundance of STAT3, SHP2, and RANKL in SHP2-sufficient and SHP2-deficient OBs in the presence or absence of the STAT3 inhibitor C188. In all western blotting studies, ACTIN served as an internal loading control. The low-power full images of Fig. 4a–c shown in Figs S2c and S4b