Cell Lysis, Immunoblotting, and Migration Assay

Cell lysis and immunoblotting were performed as described previously (34 (link)). For phosphatase treatment, cell lysates were incubated with alkaline phosphatase (E. coli C75) (TAKARA BIO INC.) in the reaction buffer (50 mM Tris-HCl (pH8.8), 1 mM MgCl₂) for 30 min at 60 °C. Actin stress fiber formation was detected by staining with Rhodamine-conjugated phalloidin (Cytoskeleton), as described in Motizuki et al. (62 (link)). Chamber migration assay was performed as described in Motizuki et al. and Lee et al. (11 , 33 (link)). Quantitative real-time PCR was performed as described in Itoh et al. (63 (link)). Primer sequences are shown in Table S1.