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. 2020 Jul 7;14:2657–2665. doi: 10.2147/DDDT.S241826

Figure 4.

Figure 4

Osteoclast differentiation analysis. (A) qRT-PCR detected gene expression of Nfatc, c-fos, RANK, and MMP9 in RAW264.7 cells co-culture with PBS (ctrl), CpG ODN (ODN), phosphorothioate CpG ODN, and N-Ac-L-Leu-PEI/CpG ODN complexes (N-Ac-L-Leu-PEI/ODN, w/w= 4:1) for 4 days. Data represents the mean ± SD of three independent experiments and are statistically analyzed with the Student’s t-test. *P<0.05, **P<0.01, ***P<0.01 vs the ctrl group; #P < 0.05, ##P <0.01 vs phosphorothioate ODN group. (B) Western blot detected protein expression of Nfatc, c-fos, RANK, and MMP9 after co-culture with PBS (ctrl), CpG ODN (ODN), phosphorothioate CpG ODN, and N-Ac-L-Leu-PEI/CpG ODN complexes (N-Ac-L-Leu-PEI/ODN, w/w= 4:1) for 4 days. (C) The TRAP+, F-actin ring, and multinuclear cells were examined after co-culture with PBS (ctrl), CpG ODN (ODN), phosphorothioate CpG ODN, and N-Ac-L-Leu-PEI/CpG ODN complexes (N-Ac-L-Leu-PEI/ODN, w/w= 4:1) for 4.5 days by TRAP staining, rhodamine phalloidin staining, and Hoechst staining, respectively. Data represents the mean ± SD of three independent experiments.