Research Paper Volume 6, Issue 4 pp 248—263
Methylated TRF2 associates with the nuclear matrix and serves as a potential biomarker for cellular senescence
Figure 1. Methylated TRF2 is associated with the nuclear matrix. (A) Sequential extraction of the nuclear matrix from hTERT-BJ cells. Immunoblotting was performed with anti-TRF2-2meR17, anti-TRF2, anti-hRap1, anti-TRF1, anti-Lamin A, anti-H2AX, or anti-PRMT1 antibody. (B) Western analysis of 293T cells expressing shPRMT1 or the vector alone. Immunoblotting was performed with anti-PRMT1, anti-TRF2-2meR17 or anti-TRF2 antibody. The γ-tubulin blot was used a loading control. (C) Western analysis of 293T cells overexpressing Myc-tagged wild type TRF2, TRF2 carrying amino acid substitutions of arginines to lysines (TRF2-RK) or TRF2 lacking the N-terminal GAR/basic domain (TRF2-ΔB). Immunoblotting was carried out with anti-TRF2-2meR17 or anti-Myc antibody. (D) Sequential extraction of the nuclear matrix from IMR90 cells. Immunoblotting was performed with anti-TRF2-2meR17, anti-TRF2, anti-Lamin A or anti-H2AX antibody. (E) Sequential extraction of the nuclear matrix from GM9503 cells. Immunoblotting was performed with anti-TRF2-2meR17, anti-TRF2 or anti-Lamin A antibody. (F) Sequential extraction of the nuclear matrix from HeLaI.2.11 cells. Immunoblotting was performed with anti-TRF2-2meR17, anti-TRF2, anti-Lamin A or anti-H2AX antibody. (G) Western analysis of early and late passage GM9503 cells. Immunoblotting was performed with anti-TRF2-2meR17 and anti-TRF2 antibody. The γ-tubulin blot was used as a loading control.