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

Figure 1. Sir2p function is dependent on Hsp82.

Figure 1

(Ai) The temperature sensitive strain iGD170hsp82 causes disruption of mating type silencing at restrictive temperature (37°C), where Hsp82 is non functional. (Aii) iG170Dhsp82 at 37°C shows considerable amount of HMLα2 transcript comparable to that present in Δsir2 cells; whereas at 25°C negligible amount of HMLα2 is seen. (Bi) Semi quantitative RT-PCR shows increase in YFR057w transcript in iG170Dhsp82 at 37°C compared to 25°C, indicating that the strain has defect in telomere position effect, whereas wild type strain (HSP82) shows silencing at higher temperature (37°C). (Bii) Graphical representation of four independent experiments of (Bi) is done after normalization with ACT1 control. (C) Western blot shows the relative abundance of Hsp82 and Sir2p at two temperatures. Actin is the loading control. (D) The quantification of Western blot (from 4 independent experiments) shows more than 50% reduction in steady state level of Sir2p at higher temperature. The data are normalized with respect to the loading control Actin. Each bar represents mean density ± SD.