Fig. 6: Ectopic expression of nematode ANOH-1 or human ANO1 confers mechanosensitivity to ASK neurons.

a MRCs of wild-type ASK neurons or ASK neurons ectopically expressing nematode ANOH-1 or human ANO1. Left: sample traces; Right: peak MRC amplitudes. Mechanical stimulation: 15 μm displacement. Holding potential: −70 mV. P values were calculated using Kruskal-Wallis test. b MRCs in ASK ectopically expressing ANOH-1. Left: sample traces; Right: peak MRC amplitudes. P values were calculated using Kruskal-Wallis test. c MRCs recording in ASK neurons ectopically expressing ANOH-1. Left: representative traces of MRCs. The cell membrane was initially voltage-clamped at −50 mV, 0 mV, and 50 mV, and the displayed voltages were subsequently corrected for liquid junction potentials (LJPs). Middle: I-V relationship of MRCs. Right: the reversal potential of MRCs. Mechanical stimulation: 15 μm displacement. P values were calculated using Brown-Forsythe and Welch ANOVA tests. Mutations in the predicted pore region of ANOH-1 abrogated mechanosensitivity of ASK neurons with ectopic expressing of ANOH-1. d Sequence alignment of the putative pore region of ANOH-1 and its homologs. The positively charged residues K and R between TM5-TM6 regions are highly conserved among ANOH-1 and its homologs. Homology alignment was performed using the MEGA11. e No MRC was recorded in ASK neurons ectopically expressing ANOH-1 with either K588E or K588A mutations. Left: sample traces. Right: peak MRC amplitudes. Mechanical stimulation: 15 µm displacement. Holding potential: −70 mV. f Membrane topology of ANOH-1 and the location of positively charged K588 residue. P values were calculated using Kruskal-Wallis test. Day 2 adult hermaphroditic animals were used in these experiments. Each dot represents 1 animal. Data are presented as mean ± SEM. ns not significant, **P < 0.01, ***P < 0.001, ****P < 0.0001. Source data are provided as a Source Data file.