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. Author manuscript; available in PMC: 2012 Dec 1.
Published in final edited form as: Int J Biochem Cell Biol. 2011 Sep 3;43(12):1792–1801. doi: 10.1016/j.biocel.2011.08.018

Fig. 4.

Fig. 4

Effect of the purified P1α/P2β heterodimer on the depurination activity of Stx1A1 and Stx2A1 on yeast ribosomes. Yeast ribosomes (20 pmol) from the wild type (A), ΔP1 (B) and P0ΔAB (ΔAB) mutants (C) were incubated with 0.3 pmol of activated Stx1 (black bars) or Stx2 (open bars) in the presence of different amounts of P1α/P2β dimer in a final volume of 100 μl at 30°C for 30 min. The rRNA was isolated and 500 ng was used to quantify the relative level of depurination using qRT-PCR by the comparative ΔCT method (ΔΔCT). The y-axis shows the fold change of toxin-treated samples over the control samples without toxin treatment (No toxin control, NC). RIP:P1α/P2β ratios were 1:10, 1:100 and 1:1,000, which corresponded to ribosome:P1α/P2β ratios of approximately 1:0.15, 1:1.5 and 1:15 as indicated on the x-axis. The analysis was repeated three times using different preparations of ribosomes.