Skip to main content
. 2015 Oct 28;6:8759. doi: 10.1038/ncomms9759

Figure 1. Furins cleave Trunk intracellularly.

Figure 1

(a) Trk:Ch consists of the Trk coding sequence fused to three Myc epitopes and the fluorescent protein mCherry (Ch). NTrk:Ch is Trk:Ch with the cysteine-knot region removed. SP, signal peptide. (b) Immunoblot (anti-Myc) of Trk:Ch expression in embryos and S2 cells. Full-length Trk:Ch (predicted to be 55 kDa, arrowed), and three cleaved Trk:Ch species (1–3) are detectable in embryos (Emb) and both the S2 cell pellet (P) and supernatant (SN). (c) Immunoblot of NTrk:Ch expression in S2 cells. Full-length (predicted to be 44 kDa, arrowed) and cleaved (1) NTrk:Ch are detectable in the cell pellet and supernatant. (d) Immunoblot of supernatant from Trk:Ch-expressing S2 cells incubated with (+) and without (−) Furin inhibitors over 3 days. Trk:Ch cleavage is markedly reduced or absent when Furin inhibitor is present (compare species 1 level in lanes with and without inhibitor). For uncropped immunoblot see Supplementary Fig. 1a. (e) Immunoblot and quantification of NTrk:Ch in S2 cell supernatant. Cleavage is significantly reduced when Furin inhibitor is present (unpaired t-test, *P<0.05, **P<0.01). For uncropped immunoblot see Supplementary Fig. 1b. Error bars represent ±1 s.e. n=3 for each mean. Images are representative of at least three replicate immunoblots.