Fig. 7: CIB proteins are required for the mechanosensory function of ANOH-1/ANO1.

a The touch-evoked calcium increases in CEM. Each dot represents 1 animal. P values were calculated using Kruskal-Wallis test. b Nose touch induced reversals of Day 2 adult male animals. Each dot represents 1 independent assays, and 10 ~ 15 animals were used for each assay. P values were calculated using Brown-Forsythe and Welch ANOVA tests. c The touch-evoked calcium increases in ASJ. Expression of either nematode CALM-1 or human CIB2, but not human CIB3, in ASJ neurons was able to rescue the defects observed in the calm-1 mutants. Left: calcium responses; Right: maximum ΔF/F₀ changes. Mechanical stimulation: 15 μm displacement. Each dot represents 1 animal. P values were calculated using Kruskal-Wallis test. d MRCs in ASJ. Left: calcium responses; Right: maximum ΔF/F₀ changes. Each dot represents 1 animal. P values were calculated using Kruskal-Wallis test. e MRCs were detected in the ASK neurons of calm-1 mutant worms that ectopically expressed nematode ANOH-1 with CALM-1 or human ANO1 with CIB2. Left: sample traces; Right: peak MRC amplitudes. Each dot represents 1 animal. P values were calculated using Kruskal-Wallis test. f Co-IP experiments with HEK293T cells showing the direct binding of exogenously expressed nematode ANOH-1 with CALM-1, or human ANO1 with CIB2. Repeated for twice. g ANOH-1/ANO1 acts as a core component of a mechanosensory anion channel complex. The CIB and ankyrin proteins are likely auxiliary components of the mechanotransduction channel complex. Ectopic expression of ANOH-1 or human ANO1 channels confers mechanosensitivity to mechanically insensitive neurons through a Calm-1/CIB2-dependent mechanism, suggesting that the role of Anoctamin 1 in mechanotransduction is evolutionarily conserved. Data are presented as mean ± SEM. ns: not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Source data are provided as a Source Data file.