Research Paper Volume 4, Issue 8 pp 567—577
A filtering strategy identifies FOXQ1 as a potential effector of lamin A dysfunction
Figure 2. Ectopic expression of FOXQ1 in normal human diploid fibroblasts results in reduced growth rates and altered nuclear membrane morphology, two phenotypes that are observed in cells expressing progerin or elevated levels of prelamin A.
(A) Human diploid fibroblasts expressing flag-tagged FOXQ1, progerin or prelamin A were lysed and analyzed by immunoblotting with flag antibodies. Antibodies against glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were used as loading control. (B) Normal human diploid fibroblasts were transduced with lentiviruses for the expression of flag-tagged FOXQ1, progerin or prelamin A and cell growth was monitored over several passages. Control cell line represents human diploid fibroblasts transduced with a lentivirus for the expression of GFP. (C) Percentage of cells with dysmorphic nuclei at passage 3 and 4 in each experimental sample was determined as described in the materials and methods section. (D) Representative images DAPI stained nuclei in control cells and cells overexpressing progerin or FOXQ1 showing altered nuclear morphology. Arrows point to examples of nuclear blebbing.