Niclosamide potentiates endogenous CaCC in murine aortic vascular SMCs. (A) Representative whole-cell CaCC current recorded from the aortic SMCs in response to 5 µM niclosamide (Niclo) and 10 µM Ani9. The intracellular solution contained 0.5 µM free Ca²⁺. Voltage protocol is shown on the left. (B) Representative current traces induced by a voltage-step protocol from −100 to +140 mV with 20-mV increments and a holding potential at −60 mV. (C)G-V relationship of the current recorded in B. The tail currents were normalized to the tail current elicited by a +140-mV voltage step before niclosamide (G_Basal). (D) Comparison of normalized tail current elicited by a +140-mV voltage step with and without niclosamide. Statistical analysis was done by one-way ANOVA followed by Tukey’s multiple comparisons test. **: P < 0.01, n = 5. ns: not significant. (E) Representative GCaMP8.1 images of intracellular Ca²⁺ changes in response to 5 μM niclosamide and 10 μM Ani9, a specific TMEM16A inhibitor. The aortic SMCs were isolated from the Acta2-GCaMP8.1/mVermilion mice. (F) Time-course of Ca²⁺ dynamics in response to niclosamide and Ani9. Error bars represent the SEM of 23 cells from four mice. (G) Normalized Ca²⁺ signal (to basal level) after application of niclosamide and Ani9. Statistical analysis was done using one-way ANOVA followed by Tukey’s multiple comparisons test. ***: P < 0.001, n = 23 from four mice.