Effect of RRP45 depletion on mRNA abundance and stability. Trypanosomes expressing different reporter RNAs (indicated above the graphs) were treated with actinomycin D, and RNA was isolated at the various times thereafter. RNAs were detected by Northern blotting and hybridization with [32P]-labeled probes, and quantitated by phosphorImager using the SRP RNA as an internal control. For each cell line, we show a control Western blot showing typical RRP45 depletion for the cell line. Beneath the graphs are typical Northern blots. Point 0 is before actinomycin D addition; the first time point corresponds to cells that were centrifuged immediately after actinomycin D addition. (Processing takes 5–10 min.) As for Figure 1, the plus sign (+) indicates that normal levels of RRP45 are present and the vertical, downward-pointing arrow indicates that the level of RRP45 is reduced. Results from three or more experiments are displayed as mean and standard deviation.
