Fig. 3

Neither SERCA2 co-expression nor the linker-mutations affect the expression of Piezo1 in plasma membrane. a and e, Live immunofluorescent staining of the extracellularly localized Flag-tag inserted after the residue A2419 of the Piezo1-GFP, 2172–2181(10A)-GFP, and KKKK/AAAA-GFP fusion proteins from HEK293T cells transfected with the indicated constructs. The GFP images were taken as control for the expression of the fusion proteins. Scale bar, 5 μm. b and f, Scatter plots of the fluorescence intensity ratio of the anti-Flag signal (F568) over GFP signal (F488). Each dot represents the ratio of F568/F488 from an individual cell. One-way ANOVA with multiple comparison test. c and g, Western blots of the biotinylated or whole-cell lysate samples derived from HEK293T cells transfected with the indicated constructs. d and h, Scatter plots of the normalized biotinylated Piezo1 levels of cells transfected with the indicated constructs. Unpaired student’s t-test (d) or One-way ANOVA with multiple comparison test (h). Data shown as mean ± s.e.m. ***p < 0.001