Figure 6

Lifeact-mTurquoise2 protein can be used to identify many different structures in adult Nematostella vectensis . A) DIC image of a juvenile polyp approximately two weeks after fertilization. B) SEM image of the oral portion of the animal showing the ciliated cells at the surface. C) Laser confocal microscopy stack image of the aboral end of the polyp labeled with anti-alpha tubulin (secondarily labeled with Alexa 594) showing the tubulin that is present in ciliated cells. D) SEM image of the body wall of a polyp showing long cilia (white arrow) surrounded by ciliary cones. E) TEM image of a single central cilium with typical 9 + 2 arrangement of microtubules and nine stereocilia surrounding it. F) Body wall of a polyp originally injected during embryogenesis with Lifeact-mTurquoise2. Lifeact-mTurquoise2 highlights the stereocilia structures (white arrow) identified in D,E. G-H) Phallacidin labeled fixed polyp showing the intricate network of F-actin in the muscular tissue within the body (G) as well as cellular boundaries (H, white arrow). I) Single image from a time series of a moving polyp exhibiting an increased level of Lifeact-mTurquoise2 at the site of contraction (white arrows, also see Additional file 8: Video S7.) (J) Lifeact-mTurquoise2 also binds to the actin found in cell boundaries in polyps, similar to phallacidin (H). K) TEM image of cnidocyte (cn) with a curved nucleus at the basal pole (nu) showing filamentous actin-like fibers around the apex of cnidocyte capsule (inset). L) Prominent Lifeact-mTurquoise2 fluorescence is present around the apical and lateral regions of the cnidocytes, suggesting F-actin is present around the perimeter (white arrows)