Giant fiber Gal4 driver (BDSC 79602), green channel shows giant fiber neurons, magenta channel shows background staining with nc82:

Giant fiber neurons on Codex BANC v626 dataset:

b1 mn driver (SS40980, also targets iii1 mn, iii3 mn):

Janelia image for SS40980:

b3 mn driver (SS48311, also targets hg1 MN, XBI002, WBI009):

Janelia image for SS48311:

b3 mn driver (SS98650):

Janelia image for SS98650:

SS45779 – Targeting b3 MN, iii3 MN, hg1 MN

Our line:

Janelia image:

IS69306 – b1 mn driver line

Janelia image:

Retro-Tango control crossed with GF-Gal4 (BDSC: 79602), presynaptic neurons in magenta, giant fiber neurons in cyan and background staining in gray

Retro-Tango x GF-Gal4 from Sorkac et. al (2023)

rut and rad expression in b1 and b3 motor neurons, I looked for cells express 6 genes (rut, rad, FoxP, VGlut, AD TF, DBD TF) at the same time above certain threshold:

Staining is a bit faint and brain quality a bit sub-optimal and no punctae can be observed in the FoxP channel, decreasing the gain did not resolve this, blocking was done for 1 hour at RT. Perhaps increasing concentration of 1st degree antibodies, especially for RFP might improve the results.

Green: GFP
Magenta: BRP

MBON02 trans-tango image from Kaun Lab. Left image shows trans-tango signal in red, right image shows overlay of all channels (green for MBON02, red for trans-tango, blue for nc82):

My MBON02 trans-tango images stained with anti-HA instead of anti-RFP. Flies generated by crossing trans-tango mkII line (BDSC: 95317) with MB399B (Brembslab stock number: 319). Left shows trans-tango signal in green, right shows overlay of al channels (green for trans-tango, blue for MBON02, red for nc82):

Janelia Flylight image of MB399B with 20XUAS-CsChrimson-mVenus trafficked in attP18 reporter:

Staining done by anti-GFP (MBON02), anti-RFP (post-synaptic cells), nc82 (for background stainingm not shown here). MBON02 cells are clearly visible on green channel but there is no clear cell signal on red channel (MBON02 in green channel, postsynaptic cells in red channel).

Here is the trans-tango image of MBON02 (postsynaptic cells of MBON02) from Kaun Lab:

Retro-Tango of b3 mns look worse, nearly no cell signal both in green (b3 mn) and red channels (b3 mn presynaptic neurons)