Up-regulation of S100A16 increased Notch1, ZEB1 and ZEB 2 expression. (A, B) qRT-PCR (A) and western blot (B) analyses showed that mRNA and protein levels of Notch 1, ZEB1 and ZEB2 were significantly enhanced in MCF7-S100A16 cells compared with control cells (Bars, mean ± SD, P < 0.05). (C, D) Immunofluorescence staining was used to examine the location of two transcription factors ZEB1 and ZEB2. After fixation, the celluar location of ZEB1 (red) and ZEB2 (red) were analyzed by confocal microscopy. Cell nuclei were stained with DAPI (4′, 6-diamidino-2-phenylindole, blue). Immunofluorescence staining showed that two transcription factors ZEB1 (C) and ZEB2 (D) localized in the nucleus and the fluorescence signals were enhanced after S100A16 up-regulation.