PC9 was transfected with siRNA targeting human CD44V9 (sense, 5′-CUACUUUACUGGAAGGUUAtt-3′; antisense, 5′-UAACCUUCCAGUAAAGUAGtt-3′), which has been reported previously (37) or control siRNA (sense, 5′-UUCUCCGAACGUGUCACGUtt-3′; antisense, 5′-ACGUGACACGUUCGGAGAAtt-3′) by Lipofectamine 2000 (Thermo Fisher, no. 11668019) 48 hours before performing experiment.

PIEZO1-GFP plasmid used for overexpression validation was a gift from D. Beech, which was described previously (59). PIEZO1-GFP was transfected into HUVEC cells by Lipofectamine 2000. Cells were fixed and stained with anti-PIEZO1 antibody following IF procedure described above.

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