Fig. 5: Loss of PIEZO1 obviates emergence of stem cell-like phenotypes after microcapillary-induced deformation.

A Plot of deformation index (DI) for melanoma cells transiting the microfluidic device (n = 100–150 cells per condition). c B Cell viability expressed as the mean ± SEM from three independent experiments. Statistical significance assessed with two-sided unpaired t-test: p  = 0.0001 (***) 95% CI: [-42.01, -28.66]. C, D Immunofluorescence images of melanoma cells transiting constrictive channels and quantification of fold change intracellular calcium concentration (n = 7–40 cells per condition). The results are expressed as the mean ± SEM from 2 independent experiments. Statistical significance assessed using two-sided unpaired t-test: 10 µm p = 0.0038 (**) 95% CI: [0.03575 to 0.1028]; 5 µm p = 0.0459 (*) 95% CI: [0.0008904 to 0.06797]. E, F Immunofluorescence images and quantification of nuclei with results expressed as the mean ± SEM from three independent experiments. Statistical significance assessed using a two-sided unpaired t-test: p  =  0.97 (ns), 95% CI: [−26.46, 27.12. G Immunofluorescence images of stemness markers in melanoma cells and H quantification of fold change relative to control, expressed as the mean ± SEM from five independent experiments. Statistical significance assessed using two-sided unpaired t-tests: CD44 p  =  0.0079 (**), 95% CI: [−0.7189, −0.1491]; CD271 p  =  0.037 (*), 95% CI: [−1.081, −0.04346]; ABCB5 p  =  0.0135(*), 95% CI: [−1.709, −0.2568]. I, L,M Representative images of melanoma cells forming tumorsphere after 7 days of culture, demonstration of change in tumorsphere area over time, and quantification of tumorsphere area at day 7, expressed as the mean ± SEM from four independent experiments. Statistical significance assessed using two-sided unpaired t-test: WT SQZD vs. WT SQZD_RR p < 0.0001 (****), 95% CI: [2130, 093]; WT SQZD vs. KO SQZD_RR p < 0.0001 (****), 95% CI: [2051, 5013].