Fig. 7. Locomotor and postural defects in ric-8 S467A or S472A C. elegans mutants.

(A to D) WT (N2) and CRISPR-modified C. elegans strains expressing ric-8 with alanine mutations at sites Ser⁴⁶⁷ or Ser⁴⁷² were treated with 10 μM phorbol ester or ethanol (vehicle) for 60 min. (A) Representative images show the rod-like body posture phenotype of S467A and S472A mutants on vehicle plates, as compared to the sinusoidal body posture of N2 controls. Phorbol ester–exposed animals had a hyperflexive phenotypes in N2 controls and S467A and S472A mutants, denoted by the omega-shaped body postures. Red arrowheads denote eggs laid by ric-8 S467A animals in response to phorbol ester. Scale bar, 0.5 mm. (B) The numbers of body bends were quantified in freely moving animals over a 2-min period. (C) The maximum bending amplitude was quantified as a measure of hyperflexion. (D) Track length (forward + reverse movement) was quantified in freely moving animals for 2 min before and after 60-min phorbol ester treatment using WormLab acquisition and analysis software. For all experiments, ten 1-day-old adults were assayed per plate with 10 replicated plates per strain for all experiments. Statistical significance was determined by one-way ANOVA with Tukey’s post hoc test. **P < 0.005, ***P < 0.0001.