Additional parameters calculated from ICCS for assessment of PIEZO1-GFP/CtxB–Alexa Fluor 555 colocalization in the plasma membrane. (A) Fraction of total PIEZO1-GFP colocalized with CtxB–Alexa Fluor 555. (B) Average number of CtxB–Alexa Fluor 555 entities colocalized with PIEZO1-GFP per square micrometer. (C) 2D effective diffusion of CtxB–Alexa Fluor 555 in the basal plasma membrane. One-way ANOVA; ****, P < 0.0001; error bars represent SEM.