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. 2023 May 11;12:e85041. doi: 10.7554/eLife.85041

Figure 4. Comparison of the retro-Tango signal with the EM reconstruction of the female hemibrain.

(A) Plotting of the skeletonizations of the EM segmentations of presynaptic partners that connect with the GF via 17 synapses or more. (B) Presynaptic partners of the GFs in a female fly as revealed by retro-Tango. 15do females heterozygous for the tdTomato reporter were analyzed for panel (B). Presynaptic mtdTomato (magenta) and neuropil (grey). Scale bar, 50 μm. Note the high similarity between the patterns in both panels.

Figure 4.

Figure 4—figure supplement 1. Methodology for the comparison of retro-Tango results with the hemibrain connectome.

Figure 4—figure supplement 1.

(A) Flowchart explaining the steps in the comparison. (B) Driving retro-Tango from the GFs results in nuclear staining in an average of 191 neurons in ten hemibrains. (C) In the absence of a Gal4 driver, retro-Tango has background nuclear staining in 26 neurons. The areas analyzed are marked in light grey based on the approximate regions covered by the published hemibrain connectome. 15do females heterozygous for the nls-DsRed reporter were analyzed for panels (B) and (C). Presynaptic DsRed (magenta) and neuropil (grey). Scale bars, 50 μm.
Figure 4—figure supplement 2. retro-Tango reveals LPLC2s as presynaptic partners of the GF in females when the reporter is homozygous.

Figure 4—figure supplement 2.

Initiating retro-Tango from the GFs in females homozygous for the reporter results in presynaptic signal in LPLC2 (arrow) neurons (157±20 neurons in 5 brains). 15do females homozygous for the tdTomato reporter were analyzed. Presynaptic mtdTomato (magenta) and neuropil (grey). Scale bar, 50 μm.