Extended Data Fig. 1: Characterization of the high-throughput cellular stretch system and the cellular response to mechanical strain.

a, Image of the HT cellular stretch system. b, Finite element analysis of the PDMS membrane under different vacuum levels, showing the strain pattern across the whole well. c, Illustration of the particle imagery assay used to empirically estimate the strain values with the formula shown. d is the distance of the bead from the centre of the well at the resting state, and d’ is the distance of the same bead from the centre in the stretched state. d, Bead images before stretch (grey) and at −30 kPa (magenta) were overlayed. e, Estimated strain values at different vacuum levels from the bead imagery analysis (n = 10 wells from 3 independent experiments). f, YFP intensity traces of HeLa cells subjected to 5 s stretch at a series of strain levels. Each trace is from 150 ~ 200 cells, n = 3 ~ 4 trials per group. g, The inhibition curve of the DCPIB, a non-specific chloride channel inhibitor, on the quenching of YFP induced by 40% strain. Each data point is from ~200 cells of each condition, n = 5 trials per group. h, Inhibition curve of the DCPIB on the quenching induced by 50% strain for 5 s. Each data point is from ~220 cells of each condition, n = 5 trials per group. Half-inhibition concentrations (IC₅₀) are indicated. i, YFP quenching over the course of 60 min. Fluorescence baseline was recorded for 10 min, then the following stimulations were applied: 5 µM RSL3, TSZ, and 500 ng ml⁻¹ LLO. Mechanical stimulation was applied for 5 s at 50% and 55% strain. Control was 0.5% DMSO. Each data point is from ~150 cells of each condition, n = 3 ~ 4 trials per group. j, YFP quenching over the course of 60 min under 20 ng ml⁻¹ nigercin or mechanical strain at 50% or 55% for 5 s. Each data point is from ~150 cells of each condition, n = 3 ~ 4 trials per group. Unless otherwise noted, all data are presented as mean ± s.e.m.

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