Fig 1. Transcriptome-wide polysome profiling upon heat shock of procyclic forms.

(A) Representative polysome profiling of procyclic cells grown at 27°C (normal culture conditions) or heat-shocked for 1h at 39°C. Fractions were pooled as described and used for the analysis (F: free, S: ribosomal subunits, L: monosomes and light polysomes, H: heavy polysomes). (B) For normalization, RNA prepared from four pools was analysed by Northern blotting. The total signal from the spliced leader RNA (present at the 5’-end of each trypanosomal mRNA) was used to calculate the distribution of total mRNA in four pools (mean, n = 2). The strong spot below 200nt is the spliced leader precursor RNA (SLRNA) and the smear above is trans spliced mRNA. (C) Quantitation of Northerns as in B, showing the percentage of total signal in each sucrose gradient fraction (2 biological replicates used for sequencing).