In vivo localization of NvaPKC::mVenus during different stages of N. vectensis development. The overexpression of NvaPKC::mVenus (A) protein, by mRNA microinjection, displays similar patterns observed with the antibody staining against the same protein, and no signal was observed in the endoderm. This messenger was co-injected with Lifeact::mTq2 mRNA (B) to visualize the shape of the cells and the apical cortex. As is seen in (C), NvaPKC::mVenus is below the cortical actin during all observed stages. White arrows indicate astral/centrosomal and (pro)nuclear structures. White arrowheads indicate the locations adjacent to other cells (cell contacts) where NvaPKC::mVenus is localized. Images of unfertilized, cleavage, and side panels in gastrula stages correspond to a single slice from z-stack series. Image of the whole blastula and gastrula corresponds to a 3D reconstruction from z-stack series. Side panels in blastula and gastrula stages are a mid section from the z-stack series to show the apical distribution of NvaPKC::mVenus along the A/B axis. See Additional files 5, 6, 7, 8, and 9 for more details of every stage presented here in C.