Figure 3. Developmental time course of F48E3.8 and C54G7.3 gene expression in germline-ablated C. elegans using RT-PCR. Synchronous non-permissive (germline-ablated) worms of this strain were grown on solid media and used to collect RNA from various time-points following L1 arrest at hatching and continuing into adulthood. First strand cDNA (+) was generated from each RNA extract (including No RT controls, -) and used as a template for PCR to analyze expression of the genes F48E3.8, C54G7.3, the eggshell chitin synthase
chs-1 and the pharyngeal chitin synthase
chs-2. The gene
ama-1, encoding the large subunit of RNA polII, was used as a positive control (as done by Johnstone and Barry 1996 and Veronico et. al. 2001) although we note that expression of the gene is not consistent as previously described. Ablation of the germline was confirmed by the attenuation in expression of
chs-1. The transition to adult stages was confirmed by the expression of
col-19, an adult specific collagen gene. All primers and PCR conditions are listed in Table S2.