Extended Data Fig. 2: Testing BAcTrace in tissue culture.
From: BAcTrace, a tool for retrograde tracing of neuronal circuits in Drosophila
a, Western blot analysis of S2 cell extracts transfected and incubated with decreasing amounts of toxin. b, Constructs used in (C) and (F). c, Immunochemistry of S2 cells transfected with sensors and TEV variants. d, Prediction of N-glycosylation sites on TEV (http://www.cbs.dtu.dk/services/NetNGlyc/). e, TEV structure (PDB id 1lvb)53 with potential glycosylation sites from (D) highlighted in blue. Bound pseudo-substrate shown in yellow. f, Western blot analysis of wild type and TEVT173V ability to cleave their target sequence. * non-specific band. g, Western blot analysis of cleavage efficiency under varying concentrations of hTfR::GFP receptor. Epitopes detected by antibodies are indicated in bold in (A), (C), (F) and (G). Experiments in (A), (F) and (G) were done once. Empty arrowheads in (A), (C), (F) and (G) indicate uncleaved while solid arrowheads indicate cleaved products.
