Western blot analysis of translation initiation factors. Equivalent amounts of total protein (75 μg) extracted from wild type (wt) and myo1Δ strain cultures were analyzed for the steady state levels of A) eIF2αp and it's phosphorylated form eIF2α-P, B) eIF4Gp, C) eIF4Ep. Shown are equivalent autoradiography exposures of each membrane probed with their corresponding antibodies. Pgk1p was used as a loading control in each experiment while unphosphorylated eIF2αp served as an additional loading control in panel A. The histogram below each panel illustrates the ratio of the intensity from each test protein band relative to the intensity of its PGK loading control, averaged from duplicate experiments. See Methods for details.
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