Figures Legend:
FIGURE 1 Interaction of lamin A and trf2 in the connection with telomere. A, Bioinformatic approach of finding the interaction between lamin A and telomeric repetitive sequences. Represents interaction between Lamin A (from 607 to 646 amino acids) with telomere DNA using DP-Bind bioinformatics tool. B, Represents interaction between trf2 (from 509 to 542 amino acids) with telomere DNA using DP-Bind bioinformatics tool. C, DNA-Protein interaction assay. Represents fluorescent intensity of Lamin A and trf2 interaction with telomeric repetitive DNA sequences using DNA binding assay. D, Immunoprecipitation assay. Lane 1 – Protein ladder; Lane 2 to 8 – immunoprecipitated samples stained with anti-lamin A antibodies. E, Western blot validation of trf2 and Lamin A co-immunoprecipitation with anti-Lamin A and anti-trf2 antibodies. Lane 1, co-immunoprecipitation of Lamin A protein with anti-Lamin A antibody. Lane 2, co-immunoprecipitation of Lamin A protein with anti-trf2 antibody.
FIGURE 2 Immunofluorescence assay and nuclear structural integrity of wild type and HGPS mutant lamin A. Cells were stained with anti-lamin A antibody and counter stained with DAPI. Arrow mark indicates the folding and wrinkles in nuclear membrane.
FIGURE 3 Schematic representation and overview of wild type / HGPS mutant lamin A and trf2 interaction in nuclear structural integrity.
FIGURE 4 Transfection of wildtype and mutant lamin A and functional studies. A, Immunoblotting analysis of wildtype and HGPS mutant lamin A transfected fibroblast cells stained with anti-lamin A antibodies, Lane 1 – Protein marker, Lane 2 – Wild type lamin A transfected sample, Lane 3 & 4 – HGPS mutant lamin A transfected sample. Black arrow mark indicates wild type lamin A, whereas red arrow mark indicates HGPS mutant lamin A. B, Analysis of wild type and HGPS mutant lamin A blot using Gel analyzer software. Black and red asterisk indicates wild type and HGPS mutant lamin A, respectively. C, Chromosomal fusion assay – Control. D, Chromosomal fusion analysis of HGPS lamin A mutant.