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. 2011 Oct 20;2(10):e221. doi: 10.1038/cddis.2011.104

Figure 3.

Figure 3

Hsp27 regulates eIF4E and mediates cytoprotection. (a) Western blot analysis of Hsp27, eIF4E and GAPDH protein levels in MiaPaCa-2 cells treated with OGX-427 or ASO control. (b) Histograms of average densitometries of eIF4E protein level after normalization to GAPDH protein level by densitometry analysis in MiaPaCa-2 cells after treatment with OGX-427 or ASO control. (c) Western blot analysis of Hsp27, eIF4E and GAPDH protein levels in MiaPaCa-2 cells stably transfected with empty vector (MiaPaCa-2-Mock) or human Hsp27 (MiaPaCa-2-Hsp27). (d) Western blot analysis of Hsp27, ubiquitin and eIF4E protein levels after eIF4E immunoprecipitation (IP) using rabbit anti-eIF4E antibody (reIF4E) in MiaPaCa-2-Mock and MiaPaCa-2-Hsp27 cells. Total cell lysate (TCL) represents proteins from MiaPaCa-2-Mock vs MiaPaCa-2-Hsp27 cells, extracted from cultured cells and blotted as control with Hsp27 and GAPDH antibody. (e) Histograms of average densitometries of ubiquitine protein levels after normalization to eIF4E protein levels by densitometry analysis in MiaPaCa-2 stably transfected with Hsp27 or Mock control. (f) MTT quantification of cell viability of MiaPaCa-2-Mock and MiaPaCa-2-Hsp27 cells treated with 20 nM control- or eIF4E-siRNA and gemcitabine (150 mM). Error bars represent S.E. from three independent experiments. Statistical analysis used t test; ***P≤0.001