Figure 1. Dauer formation in C. elegans and P. pacificus following Dafadine treatment: (A) C. elegans adult cultured on DMSO control plate. (B) Adult C. elegans cultured on 25 µM Dafadine exhibits characteristic protruding vulva (Pvl) and defect in the migration of distal tip cells in the gonad (Mig) phenotypes, in addition to the Daf-c dauer larvae (DL) phenotype. White arrows indicate the distal tip cell location. Scale bar represents 50 µm. (C-D) Compared to P. pacificus adults cultured on DMSO, adults on 25 µM Dafadine do not show Mig or Pvl phenotypes. (E) Synchronized eggs on DMSO control or Dafadine plates were scored for Daf-c DL and Mig phenotypes on Day 4. (F) Dafadine also did not enhance the J2 pre-dauer (J2d) or Daf-c dauer larvae (DL) phenotype of
Ppa-hsd-2(
csu60) on Day 5. P>0.05 Dunnett's multiple comparisons test to 0 µM Dafadine. (G) The developmental rate of wild-type P. pacificus shown in (E) was noticeably delayed at the highest concentration of Dafadine. *P<0.05 Dunnett's multiple comparisons test to 0 µM Dafadine. Error bars indicate the standard error of the mean. A minimum of 3 assays were performed per condition. (H) A working model of dafachronic acid production from dietary cholesterol in P. pacificus.