Figure 4. pnc-1 animals have a reduced response to the acetylcholine agonist levamisole.

(A) Spicule protractor muscles do not respond to levamisole. The percentage of males with prolonged spicule protraction (>10 sec) in response to levamisole at the indicated concentration is plotted. Spicule protraction was scored for 16–132 males at each concentration and five to seven concentrations were tested for each strain/condition. Solid lines indicate normal culture conditions, with wild-type males in black and pnc-1 males in gray. Dashed lines indicate pharmacological treatment: the blue line represents wild-type males raised with 25 mM NAM, orange and yellow lines respectively representing pnc-1 and wild-type males raised with 25 mM NA. Curves were fit and the EC₅₀ and maximal responses were determined using the GraphPad Prism software. (B) Vulval muscles do not respond to levamisole. Black columns indicate the number of eggs laid within 60 minutes per hermaphrodite in response to 0.5 mM levamisole and gray columns indicate response in buffer. NAM and NA indicate animals cultured in the presence of 25 mM nicotinamide or nicotinic acid. n= number of animals examined. Bars indicate standard deviation and **: p<0.01, *: p<0.05 (calculated using Student's t-test).