FN (Sigma-Aldrich) was coated in a final concentration of 10 μg/ml on a 48-well plate at 37°C overnight. Unbound FN was removed with PBS washing. Unspecific binding sites were blocked with 1% bovine serum albumin (BSA) for 1 hour at room temperature. Consequently, sh-ITGA5 and sh-NC hPSCs were seeded (3 × 104 cells per well) and allowed to adhere to the FN-coated plates for 30 min. Unattached cells were removed by PBS washing, and adherent cells were fixed in 4% paraformaldehyde and stained with phalloidin labeled with tetramethyl rhodamine isothiocyanate (TRITC) and DAPI. Phalloidin-labeled cells were imaged and counted.

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