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. 2016 Dec 29;11(12):e0168988. doi: 10.1371/journal.pone.0168988

Fig 2. Temsirolimus ameliorates lamin status and nuclear shape in HGPS cells.

Fig 2

(A) Representative Western blots of lamin A/C, progerin, and β-actin in control and HGPS total cell extracts isolated from either mock-treated cells or cells treated with 1.0 μM Temsirolimus daily for a period of 9 days. (B) Fold-expression of lamin A, progerin, and lamin C was determined for each sample analyzed by western blotting with anti-lamin A/C antibody in panel (A) and normalized to βan -Actin (*p-value ≤ 0.05; n = 5). (C) The frequency of misshapen nuclei (dysmorphic) after 9 days of treatment with either the vehicle or 1.0 μM Temsirolimus. An average of 900 nuclei were examined for each condition, and each experiment was repeated 3 times. (D) Immunochemistry was performed on mock-treated or Temsirolimus-treated control (GMO3349C) and HGPS (HGADFN003) fibroblasts after 9 days using antibodies against the indicated proteins (lamin A/C, progerin, and lamin B1). Representative images are shown (n = 4). Scale-bar: 20 μm.