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. 2015 Dec 7;5:17732. doi: 10.1038/srep17732

Figure 6. GSK3β Ser9 phosphorylation and hnRNPK are both required for the NF-κB activation and NFTAc1 expression during the osteoclast differentiation.

Figure 6

(a) RAW264.7 cells were co-treated with NF-κB inhibitor BAY 11-7082 respectively during the first, second, and third day of RANKL treatment. TRAP staining was performed after 3 days of RANKL treatment, and the total TRAP-positive cells in each well of the 96-well microplate were counted. The data presented are the means ± SD of three wells. **/##P < 0.01. (b) Luciferase activity assays with RAW-NF-κB cells under the indicated treatments as described in material and methods section. The results were expressed as the folds of the value of non-treated control group and presented are the means ± SD of four independent experiments. **/##P < 0.01, #P < 0.05. (c) Western blotting to check the knockdown of hnRNPK in the cells used in (b). (d) Assessment of NFATc-1 mRNA level in RAW264.7 cells by real-time qPCR after the indicated treatments. The results were expressed as the folds of the value of non-treated control group and presented are the means ± SD of three independent experiments. (e) Western blotting demonstrating the effect of hnRNPK knockdown in the cells described in (d). The numbers under each protein band indicate the fold changes after normalization using GAPDH and in comparison with the control group. The results shown are representative of at least 3 experiments.