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. 2010 Nov 18;20(3):436–444. doi: 10.1093/hmg/ddq490

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Published by Oxford University Press 2010.

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Figure 2. — Assessing the farnesylation of progerin in Lmna^csmHG/+ cells. Metabolic labeling experiments with a farnesol analogue, AG, to assess farnesylation of progerin. Lmna^+/+, Lmna^HG/+ and Lmna^csmHG/+ mouse embryonic fibroblasts were incubated with AG (30 µm) in the presence (+) or absence (−) of an FTI (ABT-100, 5 µm) for 2 days. Western blots of whole cell extracts were performed with an AG-specific antibody (green) and a lamin A/C-specific antibody (red). (A) Comparison of Lmna^HG/+ and Lmna^csmHG/+ mouse embryonic fibroblasts. (B) Comparison of Lmna^+/+ and Lmna^HG/+ mouse embryonic fibroblasts. The electrophoretic mobility of progerin, lamin B1 and lamin B2 (all farnesylated proteins) are virtually identical; hence, the AG-specific antibody detects farnesylated proteins in all cell lines. However, when the A-type lamins (lamin A, lamin C and progerin) were immunoprecipitated, only progerin in Lmna^HG/+ cells was detected with the AG antibody.