Fig. 8: Oncogenic APC alters the structure and organization of key protein constituents of the Wnt condensate signaling machinery.

For in vivo STORM imaging experiments, isolated single colonocytes from PDOs were fixed and labeled with primary monoclonal rat Fzd7 or mouse LRP6 antibody fluorescently labeled with Alexa Fluor 647. A Model of the formation of Wnt proteolipid condensates in ordered plasma membrane nanodomains examined via STORM imaging. B Representative bright field and Voronoi images of isolated single colonocytes from CRC-PDOs labeled with primary anti-LRP6-AF647. Quantitative analysis of Fzd7 and LRP6 C, E cluster area, D, F cluster area relative frequency, G, H total number of receptor molecules inside clusters, I, J receptor molecule absolute density, K, L percentage of receptor molecules forming part of clustered regions, M, N total number of receptor clusters, and O, P cellular receptor cluster density in isolated single colonocytes from PDOs, respectively. Cluster area was calculated from STORM data averaged per region of interest (ROI) and the respective relative frequency was calculated from individual cluster distribution data (mean ± SD, from n = 50–2274 ROIs, exact n value is shown below each bar). Data associated with the number of single receptor molecules, receptor clusters, and their density was calculated from raw fluorescence intensity images converted to text (.txt) x–y coordinate files using Clus-Doc (mean ± SD, from n = 22–66 ROIs, exact n value is shown below each bar). Different letters indicate significant differences between WT APC (control) and mutant APC groups (experimental) (P < 0.05). Source data are provided as a Source data file.