Extended Data Fig. 1: LC-MS analysis of PORCN-mediated WNT3Ap acylation and MS analysis of pamWNT3Ap.
From: Mechanisms and inhibition of Porcupine-mediated Wnt acylation
a, The secretion pathway of WNT ligand. In the endoplasmic reticulum (yellow), WNT (purple) is modified by PORCN (orange). Transport of the modified WNT is mediated by WLS (blue). WNTs are secreted to extracellular space via vesicles. After binding to FZD (green), WNTs trigger the signal transduction. Notum (light blue) acts as a deacylase, removing the lipid of WNT to abolish the signal. The structure of palmitoleated WNT3A hairpin 2 is shown. b, Representative LC chromatograms of PORCN-mediated WNT3Ap acylation with (red) and without (blue) LGK974. ESI-MS: m/z calculated for [M+3H]3+ 840.041, found 840.039, and m/z calculated for [palmitoleated WNT3Ap+3H]3+ 918.779, found 918.778. c, Quantitative analysis of the product ratio after the reaction. Data are mean ± s.d. (n = 3 biologically independent experiments). ****P ≤ 0.0001, two-tailed unpaired t-test using GraphPad Prism 8. d, LC analysis of pamWNT3Ap. The samples (panels b and d) were applied to a HPLC column (DiscoveryBIO wide pore C5, 4.6x150mm, 5 μm) using a gradient in which percentage of solvent B increases from 20% to 80% in 6 min at 1.5 mL/min. e, Mass spectrometry analysis of [pamWNT3Ap+3H]3+. ESI-MS: m/z calculated for [M+3H]3+ (C124H196N33O28S5) 918.451, found 918.446
