(A) The secondary structure and point mutation of S100A4. H: helices. (B) DNA sequencing data of NM (E23A, D25A), CM (N65I, D67A) and CD (N87amber). (C) Protein level of S100A4, Actin and GAPDH in OECM1-parental (P) and FaDu-parental (P) was determined by immunoblot analyses. (D) Representative images of sphere formation ability of GFP, WT, NM, CM and CD expressing OECM1 and FaDu cell lines. Black arrow indicated the sphere. (*p < 0.05; **p < 0.01; ***p < 0.001) (E) Cell surface GRP78 of GFP, WT, NM, CM and CD expressing sphere cells of OECM1 and FaDu were analyzed by flow cytometry (*p < 0.05; **p < 0.01; ***p < 0.001). (F) Protein level of GRP78, Glut3 and GAPDH in sphere cells of OECM1 by immunoblot analyses. Data are represented as mean ± SD.