Fig. 2. DUSP6 was downregulated in human and experimental osteoporosis samples.

A Human normal and osteoporosis samples were collected (n = 10). The bone density of vertebral body was analyzed. The T value of human normal samples were ranged from (0.2 to 2.1, average1.01). The T value of human osteoporosis samples were ranged from (−2.6 to −5.2, average −3.3). We isolated the RNA from the vertebral body for PCR analysis to explore the expression of DUSP6. B We fed the mice until 30 months old. The tibia was collected for uCT to explore the bone loss. And the expression of DUSP6 was analyzed in the tibia from the 30-month-old and 4-month-old mice (n = 10). C Experimental osteoporosis murine model was established (n = 5). The tibia was collected from the two groups. Immunohistochemistry assay was carried out to explore the expression of DUSP6. Original scale bars: 500 μm. D The tibia was collected from the SHAM and OVX groups. Immunofluorescence assay was carried out to detect the expression and location of ACP5 (green) and DUSP6 (red). The ACP5-positive cells were regarded as an osteoclast marker. Original scale bars: 500 μm. E RANKL-induced osteoclast differentiation was carried out. We explored the expression of DUSP1 to DUSP6 during this process. Original scale bars: 200 μm. F Nucleic acid gel electrophoresis and G western blot analysis was employed to confirm the mRNA and protein level during the osteoclast differentiation (n = 3). Data in all bar graphs are expressed as mean ± SD. *P < 0.05, ^#P < 0.01.