Figure 4.

Bar, FTI and combination treatment prevent nuclear blebbing and progerin nuclear accumulation. (A,C) Representative images of western blots for lamin A/C and prelamin A. Cultures at 15% SNS were either treated with vehicle (DMSO), Bar (1 µM), FTI (0.025 µM), or combined drugs for a period of nine days. Quantification of progerin (B) and prelamin A (D). (E) Autophagy activity was determined by measuring MDC levels using fluorescence photometry. (F) Proteasome activity was determined by measuring chymotrypsin-like proteasome activity using Suc-LLVY-AMC as a substrate. (G) Representative immunofluorescence images of HGPS (HGADFN127) fibroblasts treated for nine days as indicated. Antibodies against prelamin A (green) and progerin (red) were used, and DNA was stained with DAPI. Fluorescence images were taken at 40× magnification. Scale bar: 20 µm. (H,I) The same staining as in (G) and Figure S1a was used to determine the frequency of misshapen nuclei (dysmorphic) and the number of nuclei with bright progerin signals. An average of at least 900 nuclei were counted. Graphs show mean ± SD. Representative images are shown (n = 3; * p < 0.05, ** p < 0.01).