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[Preprint]. 2023 Nov 22:2023.11.21.568166. [Version 1] doi: 10.1101/2023.11.21.568166

Fig. 3. DmCFL localizes to the postsynapse and is reduced in KD muscles.

Fig. 3.

(A) Left: Confocal images of NMJ labeled by postsynaptic membrane anti-Dlg (red) and presynaptic membrane labeled by anti-HRP (cyan). Specific DmCFL KD classes noted. Right: Dlg channel alone (grayscale). Scale bar = 10 μm. (B) Two representative confocal images of DmCFL protein at the postsynapse of the NMJ in live larvae expressing the protein trap DmCFL::GFP. Arrowheads indicate DmCFL at postsynaptic surrounding individual boutons. Scale bar = 20 μm. (C) Top: Confocal images of larval NMJ in control (left) and DmCFL KD (right) muscles labeled with anti-DmCFL (red) and anti-HRP (cyan). Bottom: DmCFL channel alone (grayscale). Scale bar = 25 μm. (D) Top: Confocal images of larval NMJ in control (left) and DmCFL KD (right) muscles labeled with p-DmCFL (red) and anti-HRP (cyan). Specific DmCFL KD classes noted. Bottom: p-DmCFL channel alone (grayscale). Scale bar = 25 μm. (E) Top: SIM images of NMJ boutons at the larval NMJ in control (left) and DmCFL KD (right) muscles labeled with anti-DmCFL (red) and anti-HRP (cyan). Bottom: DmCFL single channel (grayscale). Arrows indicate DmCFL at postsynaptic surrounding individual boutons. Scale bar =10 μm. (F) Quantification of mean postsynaptic DmCFL intensity normalized to control (control 1 ± 0.365, n = 11 NMJs; overall DmCFL KD 0.692 ± 0.217, n = 19 NMJs). (G) Quantification of mean postsynaptic p-DmCFL intensity at postsynapse normalized to control (control 1 ± 0.433, n = 24 NMJs; overall DmCFL KD 0.484 ± 0.245, n = 22 NMJs). (F) Quantifications show mean ± SD with significance calculated by student’s t test (* p < 0.05, ** p < 0.01, **** p ≤ 0.0001).