Figure 3

Effect of HE4 transfection on ANXA2 protein and gene expression. A, The immunoblot shows the expression of HE4 and ANXA2 in ovarian cells after transfection. Lanes 1, 2, 3, 4 and 5: expression of HE4 and ANXA2 before and after HE4 gene transfection in ES-2 cells; lanes 6, 7, 8, 9 and 10: expression of HE4 and ANXA2 before and after HE4 gene transfection in CaoV-3 cells; lanes 1 and 6: high HE4 expression groups after HE4 gene transfection in the ES-2 and CaoV-3 cell lines, separately; lanes 2, 4, 7 and 9: mock groups; lanes 3 and 8: untreated groups; lanes 5 and 10: HE4 low expression groups after HE4 shRNA transfection in the ES-2 and CaoV-3 cell lines, separately. Quantitative data in panel B are expressed as HE4 and ANXA2 relative to GAPDH. Real time PCR (C) results show the expression of HE4 and ANXA2 in each group. D, Immunocytochemistry images show the expression of HE4 and ANXA2 before and after HE4 gene transfection. HE4-H: HE4 high expression after transfection; HE4-L, HE4 low expression after transfection (see Additional file 1). E, Double-labeling immunofluorescence shows the colocalization of HE4 and ANXA2 in ovarian cancer CaoV-3 cells after HE4 shRNA transfection (original magnification, ×400). The nucleus (blue, A1, A2), ANXA2 (green, B1, B2), HE4 (red, C1, C2) and merged images (D1, D2) are shown.