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. 2009 Jul 1;23(13):1559–1570. doi: 10.1101/gad.524209

Figure 6.

Figure 6.

POM34 mRNA accumulates in the cytoplasm in mps2Δ 2μm-SMY2 cells. (A) Total cell extracts from wild-type and mps2Δ 2μm-SMY2 cells were fractionated into cytosolic and membrane-bound fractions. Fractions were analyzed by immunoblotting using the indicated antibodies. The lower molecular weight band (asterisk) is a cytoplasmic degradation product of Scp160 (Frey et al. 2001). POM34 and SEC61 mRNA levels were determined by quantitative RT–PCR from three independent experiments. The graph shows the ratio of cytosolic POM34 mRNA levels to the membrane-bound POM34 mRNA levels. The bars indicate standard deviation of the results from the mean value. (C) Cytosolic fraction; (M) membrane-bound fraction. (B) Subcellular localization of POM34 mRNA in the translation initiation mutant cdc33-1. Cells were grown in YPAD at 23°C and were shifted for 2 h to 37°C. The cellular fractionation and analysis was performed as in A.