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. 2017 Mar 16;7:41710. doi: 10.1038/srep41710

Figure 2. Coronin 1A was dispensable for osteoclastogenesis.

Figure 2

(a–c) BMMs were transduced with CSII-CMV-MCS-IRES2-Venus (empty vector) lentivirus or with lentivirus expressing coronin 1A, and then were cultured with M-CSF (10 ng/mL) and RANKL (50 ng/mL) to differentiate into osteoclasts. (a,b) Cultured cells were fixed and stained for tartrate-resistant acid phosphatase (TRAP). The numbers of TRAP-positive multinucleated cells (MNCs) (>3 nuclei) were counted. Data are means ± SD of three independent experiments. Scale bars, 200 μm. (Student’s t-test), NS: not-statistically significant. (c) Expression of osteoclast marker genes, Nfatc1, Dcstamp, Atp6v0d2, Acp5 and Ctsk during osteoclastogenesis. Total RNA was extracted from the cultured cells at the indicated time points and subjected to real-time PCR. (Student’s t-test), NS: not-statistically significant. (d–f) BMMs were transfected with luciferase- or coronin 1A-specific siRNA, and then were incubated with M-CSF and RANKL. (d,e), Osteoclast differentiation of the control and coronin 1A knockdown cells. The numbers of TRAP-positive MNCs (>3 nuclei) were counted. Data are means ± SD of three independent experiments. Scale bars, 200 μm. (Student’s t-test), NS: not-statistically significant. (f) Expression of osteoclast marker genes during osteoclast differentiation. Data are representative of three experiments. (Student’s t-test), NS: not-statistically significant.