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. 2000 Nov;11(11):3993–4003. doi: 10.1091/mbc.11.11.3993

Figure 4.

Figure 4

Overexpression of Int6 causes elevated Pap1-responsive gene expression without Pap1 relocalization. (A) Transformants of S. pombe h- leu1–32 (pap1+) or h- pap1::ura4+ ura4-D18 leu1–32 (pap1Δ) containing either pREP3X, or pREP3X cDNA library plasmids directing expression of full length Int6 or Int6CT, as indicated, were grown in liquid culture for 16 h at 30°C in the absence of thiamine and were processed for northern or western blotting. A northern blot was probed to detect apt1 and trr1 RNA and subsequently stripped and reprobed to detect act1 RNA as a loading control, as indicated. A western blot of whole cell extracts prepared from the same cultures was probed with antibodies directed against Pap1, p25 and Cdc2 (loading control) as indicated. (B and C) An h- leu1–32 ura4-D18 strain transformed with pREP42-GFP-Pap1 and either pREP3X (B) or pREP3X-int6 (C) was grown in minimal medium without thiamine for 24 h in the dark at 25°C. GFP-Pap1 was visualized by confocal laser microscopy. Scale bar, 10 μm.