Figure 3. ASOs lower lamin A levels and reduce the frequency of nuclear shape abnormalities in HGPS fibroblasts.

(A) Immunocytochemistry showing increased numbers of cells expressing more lamin C than lamin A after ASO E11-31 treatment. Human HGPS fibroblasts (clone 11513) were transfected once with ASO E11-31. After 5 days, the cells were processed for immunofluorescence microscopy with a lamin A antibody that binds lamin A and progerin (green) and a lamin C antibody (red). Cells transfected with a scrambled ASO served as a control. After transfection with ASO E11-31, lamin A expression was markedly reduced in many cells (arrowheads). Scale bar: 50 μm. (B) Quantitative measurements showing a higher number of cells expressing more lamin C than lamin A in ASO E11-31–treated cells. Control ASO–treated cells (white squares) expressed approximately equivalent amounts of lamin A and C. After ASO E11-31 treatment (red squares), there was an increase in the number of cells that expressed more lamin C than lamin A. Lamin C levels were scaled to the levels measured in control cells; thus, the highest levels for lamin C were limited to approximately 85 percent. (C) ASO E11-31 reduces the number of abnormally shaped nuclei in HGPS cells. Evaluation of nuclear shape after transfection with ASO E11-31. Two HGPS fibroblast cell lines (clones 1972 and 11513) were treated once with ASO E11-31 or a scrambled ASO. After 5 days, nuclear shape was assessed by two observers blinded to treatment group (n = 2 per group).