CNS selectively increases ROS production and Ca2+oscillation in mOCs. (a) Cellular pH detection during osteoclastogenesis, bar represents 200 μm. (b) Quantification of average cellular pH level. (c) Western blot analysis of ATP6v0d2 expression. (d) Oxidation kinetic experiment testing oxidative enzymatic activity of CNS at pH 4.0 and pH 7.0. (e) Intracellular ROS detection of BMMs in a time line, bar represents 200 μm. (f) Quantification of ROS positive cell number and average cellular ROS intensity. (g) Fluo-4 fluorescent images of BMMs at 72 h. Pseudo-color labeled (purple) area represents actively fluorescence ration changing cell, bar represents 400 μm. Three representative traces are presented, fluorescence ratio change recorded every 5 s for 300 s. (h) Relative mRNA expression of ATP6v0d2, NFATc1, c-FOS and western blot images of c-FOS, NFATc1 and β-actin. Images are representative of n = 3 independent experiments. The data in the figures represent the averages ± SD. Significant differences are indicated as ** (p < 0.01). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)