Fig. 4: Ano1 knock out alleviates the enhancement of mechanical unloading on osteoclast activity. | Communications Biology

Fig. 4: Ano1 knock out alleviates the enhancement of mechanical unloading on osteoclast activity.

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

Fig. 4

a QRT-PCR analysis of Ano1 mRNA level in Ano1fl/fl and Ctsk-Cre;Ano1fl/fl osteoclasts after treatment with Ctrl or MG 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 MG for 2 days (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 MG for 2 days. Scale bar, 200 μm. d Quantification of the number of multinucleated cells per cm2. (n = 130–453, from six independent experiments). e QRT-PCR analysis of NFATc1, Acp5, Ctsk and Mmp9 mRNA levels in osteoclasts Ano1fl/fl and Ctsk-Cre;Ano1fl/fl osteoclasts after treatment with Ctrl or MG. (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 MG 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.

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