Fig. 7

AKTIP localization in progerin expressing HeLa cells (A, B) Images (A) and quantification (B) of DAPI and progerin stained nuclei of HeLa cells transduced with a control vector (LV-ctr) or with progerin expressing vector (LV-progerin) and stained for progerin and DAPI. C RT Q-PCR of LMNA mRNA (including progerin) expression. In B-C mean values of two independent experiments ± SEM are shown. In A 100 nuclei were counted for each condition D 3D rendering, projections of an extended section viewed from orthogonal planes and magnified sections of LV-progerin HeLa nucleus labelled with anti-lamin B1 and anti-AKTIP antibodies. E Projections of an extended section viewed from orthogonal planes of images of LV-progerin HeLa nuclei stained with anti-lamin A and anti-AKTIP antibodies, and magnified sections. F Quantification of AKTIP foci at a distance > 0.5 µm from lamin B1 in control (LV-ctr) and progerin (LV-progerin,) HeLa cells. Each dot corresponds to a nucleus. A minimum of 15 nuclei were analyzed for each condition. G Western blotting using anti-AKTIP antibody in control (LV-ctr) and progerin expressing (LV-progerin) HeLa cells. Actin was used as loading control. H RT Q-PCR of AKTIP mRNA in control (LV-ctr) and progerin expressing (LV-progerin) HeLa cells. Mean values of two independent experiments ± SEM are shown. I Images of wild type and HGPS fibroblasts stained for lamin A/C and AKTIP showing in merge that there is no AKTIP accumulation at the site in which lamin A/C is accumulated due to progerin expression. J Images of HGPS fibroblasts stained for progerin and AKTIP and relative enlargements showing that there is no AKTIP signal at the site of progerin accumulation at nuclear rim. DNA was stained with DAPI. K Pearson correlation value of intensity plot of AKTIP-lamin A/C distribution at nuclear rim in wild type fibroblasts, and AKTIP-lamin A/C and of AKTIP-progerin distribution at nuclear rim in HGPS fibroblasts. Each dot corresponds to a nucleus. A minimum of 10 nuclei were evaluated for each condition. Mean values ± SEM are shown. *p < 0.05, ***p < 0.001, in unpaired Student t-test