Figure 2.

qPCR and immunoblotting confirms translational profiling. (A) Quantitative reverse-transcription PCR analysis of translation state for qPCR (light grey bars ±SD, n = 3) and array data (black bars) for indicated transcripts (plotted on a log₂ scale so that reduced polysome association in mutant cells is shown as a negative number). (B) Top: total protein levels of Sec9p, Taf7p, Taf3p, Cic1p, Lsm8p, Pub1p and Ade2p were analysed by immmunoblot analysis in indicated yeast strains. Experiments were done in triplicate. Loading control Arp2p (Actin Related Protein 2) is shown beneath each. Middle: densitometry analysis for each protein relative to Arp2p is beneath each panel (dark grey bars). Bottom: polysome/monosome [P/M] ratios from translational profiling (light grey bars).