Figure 4

Single nematode RT-PCR to assess transcript levels for the P-ef1b and P-rps-2 genes in control and RNAi treated Panagrolaimus sp. PS1159 nematodes. RNA was isolated from individual nematodes whose progeny showed a positive RNAi phenotype and from individual control nematodes fed on E. coli containing the empty L4440 plasmid vector. Transcript levels were determined by RT-PCR for the target gene and for the rRNA D3 gene (which served as an internal control). (A) Transcript levels for the P-ef1b and rRNA D3 genes in individual Panagrolaimus sp. PS1159 adult females fed on E. coli expressing P-ef1b dsRNA (each of the three replicates corresponds to a separate nematode); (B) Transcript levels for the P-rps-2 and rRNA D3 genes in individual Panagrolaimus sp. PS1159 females fed on E. coli expressing P-rps-2 dsRNA (each of the three replicates corresponds to a separate nematode); (C, D) Quantification of the RT-PCR products for the P-ef1b and P-rps-2 genes in control and RNAi treated Panagrolaimus sp. PS1159 nematodes. The band intensities are given in ImageQuant™ machine units (imu).