Fig 9. NEKL-associated clathrin defects are rescued by loss of AP2 activity.

(A,D) Representative confocal images of day-2 adults expressing GFP::CHC-1 within the apical region of the hyp7 epidermal syncytium. Assays were performed on NEKL-2::AID animals containing null alleles of (A–C) fcho-1(fd296) and (D–F) apa-2(fd283) in the presence and absence of auxin (20 h). Background subtraction was performed using the same parameters for all images; minimum and maximum pixel values were kept consistent for all images. Inverted fluorescence was used to aid clarity. Bar size in A = 5 μm (for A and D). (B,C,E,F) For individual adults, the mean GFP::CHC-1 intensities (B,E) and the percentage of GFP-positive pixels above threshold (C,F) were determined. (G) Fluorescence recovery curves of NEKL-2::AID fcho-1(fd296) day-2 adults in the presence and absence of auxin. Normalized average mean intensities of photobleached regions were plotted as a function of time using 5-s intervals; error bars denote SEM. (H) Bar plot showing the mobile fractions from FRAP analyses of wild-type, NEKL-2::AID, and NEKL-2::AID fcho-1(fd296) adults. (B,C,E,F) The group mean and 95% confidence intervals (error bars) are shown. (H) Error bars indicate 95% confidence intervals. p-Values were determined using two-tailed Mann-Whitney tests; **p < 0.01]. Raw data are available in S1 File.