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. Author manuscript; available in PMC: 2021 Jan 23.
Published in final edited form as: Cell. 2020 Jan 16;180(2):373–386.e15. doi: 10.1016/j.cell.2019.12.029

Figure 1. Cell-Surface Biotinylation of Olfactory Projection Neurons in Intact Brains.

Figure 1.

(A) Scheme and features of cell-surface biotinylation in intact tissues.

(B) Olfactory projection neuron (PN) specific VT033006-GAL4 (PN-GAL4, hereafter) drove the expression of membrane-targeted GFP (CD4-GFP). The zoom-in panel shows a single optical section of the PN soma area. Orange circle, antennal lobe.

(C and D) Neutravidin staining of antennal lobes after the cell-surface biotinylation reaction. (C) HRP was not expressed by omitting the GAL4 driver. (D) PN-GAL4 drove the expression of cell surface-targeted HRP (HRP-CD2). The zoom-in panel shows a single optical section of the PN soma area.

(E) Left and middle, streptavidin blots of the whole-brain lysate (left) and the post-enrichment bead eluate (middle). Right, silver stain of the post-enrichment bead eluate. −HRP, PN-GAL4 omitted; +HRP, PN>HRP-CD2.

(F) Immunoblots of intracellular proteins, actin and bruchpilot (Brp), and neuronal surface protein N-cadherin (NCad) in the whole-brain lysate and the post-enrichment bead eluate. −HRP, PN-GAL4 omitted; +HRP, PN>HRP-CD2.

Scale bar, 10 μm.

See also Figure S1.