Fig. 2. Inhibition of p43 expression by RNAi. (a) Construct used for the inhibition of p43. A map of p43 shows the 238 bp construct cloned into p4440. Bacteria were transfected with this construct and the expression of dsRNA was induced as described previously (Timmons and Fire, 1998). Euplotes were fed daily with bacteria. The position of the La motif, the RNA-binding motif (RRM with the submotifs RNP1 and 2) and primers used for PCR amplification are indicated (Aigner et al., 2000). (b) In situ staining of macronuclei with an anti-p43 antibody before (left) and after (right) RNA inhibition. (c) Western blot analysis of cells before (left) and after inhibition (right) of p43 expression. Filters were incubated simultaneously with an anti-p43 antibody and as a control with an anti--tubulin antibody (Sigma). (d) FISH analysis of telomerase RNA subunit after inhibition of p43 before (left) and after (right) electroelution. Control macronuclei (left), macronuclei after inhibition of p43 expression (right). Arrows point to the replication band. Bar, 10 µm.

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