GAGE proteins are recruited to the nuclear envelope by GCL.

A. Immunohistochemical double-staining of Myc-tagged human GCL and endogenous A-type lamins (lamins A/C) in transfected HeLa cells; GCL co-localized with lamins A/C at the nuclear envelope. B-E. HeLa cells were transiently transfected with GAGE12I (B), GCL-Myc (C), or GCL-Myc plus either GAGE12I (D) or GAGE1 (E). GCL localized at the nuclear envelope and recruited GAGE proteins from the nucleoplasm. F-G. Melanoma cell lines MZ2-MEL (F) and SK-MEL-31 (G), which express high levels of endogenous GAGE, were transfected to express GCL-Myc; immunostaining revealed GCL-mediated translocation of GAGE from the nucleoplasm to the nuclear envelope. H–K. Immunohistochemical analysis of endogenous GAGE proteins in human normal tissues and tumors revealed a dense GAGE signals near the nuclear envelope in cells of the fetal adrenal cortex (H), migrating primordial germ cells (I), breast carcinoma cells (J) and malignant melanoma cell (K) specimens. (GAGE = DAB/brown; Nuclei = Mayers hematoxylin/blue). Bars, 5 µm (A) and 10 µm (B-K).