ULK1 Suppresses Osteoclast Differentiation and Bone Resorption via Inhibiting Syk-JNK through DOK3

Figure 4

ULK1 regulates the activation of Syk/JNK through DOK3. (a) The correlation of ULK1 and docking protein 3 (DOK3) expression in osteoclast differentiation (GSE56815 dataset). (b) The expression of DOK3 in Si-NC and Si-ULK1 in BMM. (c) DOK3 expression in control and ULK1 overexpressing BMM. (d) Western blotting to detect the expression of DOK3 in Si-NC and Si-ULK1 BMM. (e) Western blotting to detect the expression of DOK3 in control and ULK1 overexpressing BMM. (f) Immunofluorescence analysis of DOK3 expression (green) in BMM between the sham and OVX groups. BMM was stained with CD11b (red). Nucleus was stained with DAPI (blue). The red arrowhead points to BMM. The white line showed the boundary between bone and bone marrow cavity (scale bar, 50 μm). (g) Immunofluorescence analysis of DOK3 expression (green) in OC between the sham and OVX groups. OC was stained with CTSK (red). Nucleus was stained with DAPI (blue). The yellow line marks OC. The white line showed the boundary between bone and bone marrow cavity (scale bar, 50 μm). (h) Expression of DOK3 during osteoclast differentiation of RAW264.7 cells. (i) Western blotting analysis of JNK signalling in Si-NC and Si-DOK3 RAW264.7 cells treated with 50 ng/ml RANKL for 0–30 minutes. (j) p-Syk, total Syk levels of Si-NC and Si-DOK3 in RAW264.7 cells. (k, l) TRAP staining (k) and quantification (l) in Si-NC and Si-DOK3 OC (scale bar, 50 μm). (m) TRAP staining and quantification in control and ULK1 overexpressing and ULK1 overexpressing OC with Si-DOK3 (scale bar, 50 μm). All data are ; .

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