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. 2021 Sep 6;4:1038. doi: 10.1038/s42003-021-02528-w

Fig. 6. N-glycosylation in the N-terminus of Piezo1.

Fig. 6

a Electrophysiological recordings of Piezo1−/− HEK293T expressing single Asn to Gln Piezo1-GFP mutations in the N-terminus (N295Q, N658Q, N885Q, N892Q, N1095Q) in the cell-attached configuration in response to negative pressure applied using a high-speed pressure-clamp (red). b Quantification of peak current elicited per patch for point mutations shown in a compared to current from Piezo1-GFP. c Representative western blot of all Asn to Gln mutants in the N-terminus of Piezo1 compared to the double mutant in the Cap region (CapQQ). d Electrophysiological recordings of HEK293T Piezo1−/− expressing Piezo1 with multiple Asn to Gln mutations (as indicated) in the cell-attached configuration in response to negative pressure applied using a high-speed pressure-clamp (red). [5N-Q—N295Q, N658Q, N885Q, N892Q, N1095Q and 7N-Q—N295Q, N658Q, N885Q, N892Q, N1095Q, N2294Q, N2331Q]. e Quantification of peak current elicited per patch for mutation combinations shown in d compared to current from Piezo1-GFP, N885Q and CapQQ. f Membrane to cytoplasmic ratio of immunogold labelling of the 7N-Q mutation compared to human Piezo1. g Representative western blot showing the comparison between Asn to Gln mutations of single and double mutations illustrating the strong impact of N885Q mutation on the upper FG band of Piezo1 heterologously expressed in Piezo1−/− HEK293T. h Representative blot showing the comparison between multiple Asn to Gln mutations 5N-Q, 7N-Q and the CapQQ (N2294Q/N2331Q). i Representative blot showing the comparison between PNGaseF digested WT and the 7N-Q mutant. *p < 0.05 determined by Kruskal–Wallis test with Dunn’s post-hoc test or Mann–Whitney U-test. −ve represents an un-transfected control.