FIGURE 2

FIGURE 2

Generating piezo1 in-frame mutant. (A) Schematic illustration of CRISPR target in piezo1 gene with a guide RNA targeting splice acceptor site of exon2. Through this guideRNA, the splice acceptor site was eliminated, generating cryptic acceptor site which deleted several bases of internal exon 2. (B) Confirmation of amino acid sequence. Quantification of relative mRNA level of (C)piezo1 and (D)piezo2a in piezo1^{11aa del/11aa del}. Values are presented as mean ± SD and analyzed using Student’s t-test. **p < 0.01. (E) Representative traces of negative pressure-induced inward currents recorded at −80 mV in negative control (NC), zebrafish Piezo1 Wildtype (WT), and zebrafish 11aa del-Piezo1. Blue arrows indicate the time when negative pressure was applied. Grey dashed lines indicate baseline of the current. (F) Measurement of current amplitude, measured in arbitrary units (AU). Values are presented as mean ± SD and analyzed using one-way ANOVA followed by Tukey’s test. ****p < 0.0001. ns indicates not significant.