A549 cells were transfected with Veh shRNA and shRNA target PIEZO1 or PIEZO2, and the stable cell line was selected and maintained in culture medium with puromycin as described in the ‘Materials and methods’ section. The mRNA expression of PIEZO1 and 2 in these cells was analyzed using RT-qPCR, and the protein expression of GAPDH, PIEZO1, and 2 in these cells was analyzed using Western blot. Cell migration in A549 cells with and without knockdown of PIEZO genes was analyzed by scratch assay as described in the ‘Materials and methods’ section. (A,E) mRNA expression of PIEZO1 (A) and PIEZO2 (E) in A549 cells. (B,F) Protein expression PIEZO1 (B) and PIEZO2 (F) in A549 cells; upper panel indicates the representative images of Western blot, and the lower panel shows the quantitation of protein expression. (C,D) Cell migration in A549 cells with stable knockdown of PIEZO1. Representative images (C) and quantitation of migration (D) in A549 cells with stable knockdown of PIEZO1. (G,H) Cell migration in A549 cells with stable knockdown of PIEZO2. Representative images (G) and quantitation of migration (H) in A549 cells with stable knockdown of PIEZO2. Data are displayed as mean ± S.D. of gene expression in different cells. *, P<0.05; **, P<0.01, cells with stable knockdown of PIEZO1 or 2 compared with cells with stable transfection of Veh shRNA.;