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. 2011 Aug 4;6(8):e23406. doi: 10.1371/journal.pone.0023406

Figure 4. Western blot analysis shows over-expression of HSP82 down regulates Sir2p in a dose dependent manner.

Figure 4

(A) Different lanes are marked with the respective genotypes. pHSP82 implies over-expression of HSP82 from a CEN plasmid. Total proteins from each lysates are probed with anti Hsp82 antibody, anti-Sir2 antibody and with anti-Actin antibody (as loading control). (B) Graphical representation of (A), where densitometric measurements of the bands from four experiments are plotted, after normalizing with Actin band intensities. Each bar represents mean density ± SD. (C) Multiple levels of Hsp82 over-expression is shown using high (2 µ) and low copy plasmids (CEN), to demonstrate dose response between Hsp82 and Sir2p levels. (D) The graphical representation of Western blot as in figure (C) shows over expression of Hsp82 from a high copy plasmid confers more reduction of Sir2p level. Each bar represents the mean density ± SD from 4 independent experiments. The data are normalized with respect to Actin.