Figure 5. Piezo1 throughout the differentiation of embryonic stem cells into motor neurons.

(A) Peak- and slow- mechanosensitive currents throughout the differentiation are plotted together with the difference between them, which we called ‘fast component’. (B) Piezo1 expression throughout the differentiation as assessed by transcriptome analysis. The expression of Piezo1 throughout the differentiation strikingly resembles the evolution of the fast component of the mechanosensitive current. FPKM: fraction per kilobase per million reads. (C) Schematic of the Crispr design. The diagram shows the beginning of the Piezo1 mRNA in mES cells. The first three exons are shown, along with the coding sequence (CDS) and the first predicted transmembrane (TM) region. Two guide RNA sequences (sgRNAs) were chosen to generate a double strand break in the beginning of the first TM region. Below the diagram, a sequence reaction of a fragment of DNA extracted from one of the modified colonies is shown. In yellow is marked the targeted sgRNA sequence, and boxed in black is marked the region with a two base-pair deletion that generates a frame-shift mutation, and an early stop codon shortly after. Only one sequence for each colony was obtained after sequencing with no background, indicating a homozygous mutation. (D) Both Piezo1 knock-out colonies of mouse embryonic stem cells showed no mechanosensitive activity.