Fig. 4: Ano1 regulates osteoclast differentiation and function via modulating intracellular chloride concentration. | Nature Communications

Fig. 4: Ano1 regulates osteoclast differentiation and function via modulating intracellular chloride concentration.

From: Anoctamin 1 controls bone resorption by coupling Cl− channel activation with RANKL-RANK signaling transduction

Fig. 4

a Representative chloride currents recorded from voltage ramps from −80 to +160 mV in whole-cell patch-clamp of RANKL-induced osteoclasts from Ano1fl/fl and Ctsk-Cre;Ano1fl/fl mice for 5 days. b Representative chloride currents recorded from voltage ramps from −80 to +160 mV in whole-cell patch-clamp of RANKL-induced osteoclasts from WT and Ano1 TG mice for 5 days. c, d Measurement of intracellular chloride concentration in RANKL-inducted osteoclasts. c Representative fluorescent images of RANKL-induced osteoclasts from Ano1fl/fl and Ctsk-Cre;Ano1fl/fl mice for 5 days (left). The relative fluorescence intensity of RANKL-induced osteoclasts (right) (n = 6 well). All the cells were stained with 5 mM MQAE. Scale bar, 10 μm. d Representative fluorescent images of RANKL-induced osteoclasts from WT and Ano1 TG mice for 5 days (left). The relative fluorescence intensity of RANKL-induced BMMs cells (right) (n = 6 well). All the cells were stained with 5 mM MQAE. Scale bar, 10 μm. e Representative images of TRAP staining in RANKL-induced osteoclasts from Ano1fl/fl and Ctsk-Cre;Ano1fl/fl mice for 5 days (left). Quantification of the number of multinucleated cells per well (right). f Representative images of TRAP staining in RANKL-induced osteoclasts from WT and Ano1 TG mice for 5 days (left). Quantification of the number of multinucleated cells per well (right). g Representative images of acridine orange staining and resorption pits in RANKL-induced osteoclasts from Ano1fl/fl and Ctsk-Cre;Ano1fl/fl mice for 5 days (left). Quantification of the resorption pit area (right). h Representative images of acridine orange staining and resorption pits in RANKL-induced osteoclasts from WT and Ano1 TG mice for 5 days (left). Quantification of the resorption pit area (right). All data are the mean ± s.e.m. from three independent experiments. Two-tailed unpaired Student’s t-test was used for statistical evaluations of two group comparisons. Source data are provided as a Source Data file.

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