Fig. 3

fbxl-1(ok3741) worms had impaired neuromotor function. (A) Pharyngeal pump rate experimental overview (Supplemental Video 1). WT C. elegans animal observed under dissecting microscope using oblique illumination; the grinder appears as a white dot (arrow) in terminal bulb. (B) Pharyngeal pumping rates of fbxl-1(ok3741) and WT worms at 1- and 6-day adult stage (n = 20 per strain at 1-day adult stage; n = 25 per strain at 6-day adult stage). Bonferroni’s correction method was applied to account for multiple comparisons, and significant findings still held. (C) fbxl-1(ok3741) and WT body bend rate on agar plates (Supplemental Videos 2 and 3). n = 25 worms per strain; 5 biological replicates performed. (D–F) Worm swimming activity analysis in liquid media. (D and E) Five worms were transferred to each drop of liquid media (scale bar: 1 mm) on a glass slide. Activity of all 5 worms in aggregate was recorded, and pixel change was analyzed using ZebraLab. (F) fbxl-1(ok3741) worm swimming activity was significantly lower than WT worms (Supplemental Videos 4 and 5). Each dot indicates the total activity of 5 worms in a single technical replicate. n = 12 for WT; n = 11 for fbxl-1(ok3741); with a total of 60 WT and 55 fbxl-1(ok3741) worms analyzed across 4 biological replicates. (G–J) Thrashing activity assay. (G) Each fbxl-1(ok3741) and WT worm was transferred to 5 μL liquid media for individual worm activity analysis by wrMTrck plugin (ImageJ). The graph shows the BBPS, which were significantly decreased in fbxl-1(ok3741) worms as compared with WT worms. Each dot indicates activity of an individual worm. n = 36 each strain. (H and I) Representative traces of worm movements detected by wrMtrck analysis. fbxl-1(ok3741) worms showed less activity and uncoordinated movements than WT worms. (J) Higher-magnification images of representative individual fbxl-1(ok3741) and WT worm traces. Data are shown as the mean ± SD. Significance was determined using unpaired Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001.