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. 2019 Apr 5;212(2):387–395. doi: 10.1534/genetics.119.302063

Figure 3.

Figure 3

lev-10::gfp11x7 yields a stronger NATF GFP signal than the single-copy lev-10::gfp knock-in at synapses in neurons and muscle cells. (A) Confocal image showing localization of LEV-10::GFP in a single-copy GFP knock-in at the native lev-10 gene (lev-10::gfp). LEV-10::GFP puncta are visible at the nerve ring (arrow), and in ventral and dorsal nerve cords (arrowheads). (B) Confocal image of the LEV-10 NATF GFP signal at body muscle synapses arising from the combination of the lev-10::gfp11x7knock-in with Pmyo-3::gfp1-10. NATF GFP (arrow) is detected at neuromuscular synapses near the nerve ring. Bar, 20 µm. Insets (right) shows rotated views of anterior regions of images on left to depict nerve ring labeling. Asterisks mark gut autofluorescence. (C) LEV-10 NATF GFP at body muscle synapses in the nerve ring labeled with lev-10::gfp11x7 is significantly brighter (around three times) (3450 ± 441) than the single-copy lev-10::gfp knock-in (1280 ± 184). P < 0.001, N = 15, Student’s t-test. Error bars are SD. (D) The LEV-10 NATF GFP signal at body muscle synapses in the nerve ring labeled with lev-10::gfp11X7 is significantly more stable (0.75 ± 0.15) to photobleaching than the single-copy lev-10::gfp knock-in in the nerve ring (0.45 ± 0.12), N = 10, P < 0.001, Student’s t-test. See Materials and Methods. Error bars are SD. AU, arbitrary units; NATF, Native And Tissue-specific Fluorescence.