Fig 2: Knockdown of ESPN inhibited ESCC cells growth in vitro. a, c The endogenous expression of ESPN in KYSE270 and TE10 cell lines transfected with ESPN-mediated siRNAs and negative control were determined at mRNA and protein levels by qRT-PCR and Western blotting. b, d The cell viabilities and colony numbers of the siESPN-ESCC and negative control-ESCC cells were measured in different time points as indicated by MTT assay and colony formation assay. Data from three independent experiments are expressed as mean ± SD

Fig 3: The cell cycle distribution, p21 and p27 expressions were determined in siESPN ESCC cells. a The cell cycle progression was analyzed by flow cytometry in negative control, ESPN-depleted KYSE270 and ESPN-depleted TE10 cells. b The protein expression levels of p21 and p27 were examined by Western blotting in negative control, ESPN-depleted KYSE270 and ESPN-depleted TE10 cells

Fig 4: Immunohistochemical staining of ESPN in esophageal squamous cell carcinoma. A Representative example of ESPN expression with no staining. Original magnification, ×200. B Representative example of ESPN expression with weak staining. Original magnification, ×200. C Representative example of ESPN expression with moderate staining. Original magnification, ×200. D Representative example of ESPN expression with strong staining. Original magnification, ×200. E ESPN immunoreactivity was present in cells in seminiferous ducts in human testis used as a positive control. Original magnification, ×200