Fig. 3: Ano1 knock out blunts the inhibition of hypergravity on osteoclast activity. | Communications Biology

Fig. 3: Ano1 knock out blunts the inhibition of hypergravity on osteoclast activity.

From: Mechanical stimulation controls osteoclast function through the regulation of Ca2+-activated Cl− channel Anoctamin 1

Fig. 3

a QRT-PCR analysis of Ano1 mRNA level in Ano1fl/fl and Ctsk-Cre;Ano1fl/fl osteoclasts after treatment with Ctrl or HG (4 g) for 2 days. (n = 3 independent experiments). b Western blot analysis of Ano1 protein level in Ano1fl/fl and Ctsk-Cre;Ano1fl/fl osteoclasts after treatment with Ctrl or HG (left). The quantification of Ano1 protein level in osteoclasts (right). (n = 3 independent experiments). c Representative images of TRAP staining in Ano1fl/fl and Ctsk-Cre;Ano1fl/fl osteoclasts after treatment with Ctrl or HG (4 g) for 2 days. Scale bar, 200 μm. d Quantification of the number of multinucleated cells per cm2. (n = 109–312, from three independent experiments). e QRT-PCR analysis of NFATc1, Acp5, Ctsk and Mmp9 mRNA levels in Ano1fl/fl and Ctsk-Cre;Ano1fl/fl osteoclasts after treatment with Ctrl or HG. (n = 3 independent experiments). f QRT-PCR analysis of NFATc1, Acp5, Ctsk and Mmp9 mRNA levels in osteoclasts isolated from Ctsk-Cre;Ano1fl/fl mice. Osteoclasts were transfected with NC, WT Ano1 or mutant Ano1 (E702/705Q) and treated with or without HG for 2 days. (n = 3 independent experiments).All data are the mean ± s.e.m. Statistical analysis with more than two groups was performed with two-way analysis of variance (ANOVA) with Šídák post-hoc test to determine group differences. *p < 0.05, **p < 0.01, ***p < 0.001.

Back to article page