Cytoplasmic capes contain ubiquitylated proteins and endosomal markers and are found in wild-type glands.

Each set of three images shows one nucleus or cape from a salivary gland showing the distribution of two proteins. The center panel is a merged red/green image with the left panel in green. Due to the varied morphology of each cape multiple examples are shown for most markers. A–P express mSPI::GFP and costaining is for the indicated proteins: A-A” – Nucleoplasmic RFP is excluded from capes (arrowheads). This image is a single plane from a nucleus where the image stack had been deconvolved; B-B” – Wheat germ agglutinin (WGA) stains glycosylated proteins of the nuclear envelope and outlines the capes indicating that they are infoldings of this membrane and that the contents are therefore extranuclear. C–F – Lamin C similarly coats the cape membranes. D,E show individual capes from other nuclei. Panel F shows a series of confocal images taken at 0.5 μm intervals through a cape at the top of a nucleus. Chambers associated with individual capes where mSpitz is very low or absent are often seen with this marker (arrowheads). G–H” – Ubiquitin (Ubi) puncta are found in most capes at their periphery (see also P–Q”). H shows a saggital view. I–J” – Occasional Hrs puncta are found in the central space of the terminal chamber; K–O – EPS15 puncta are found in most capes in the central space. L–O show individual capes from other nuclei. Panels P–U” do not express mSpi::GFP demonstrating that capes are present in wild-type glands. Costainings are as indicated: P–Q” – Ubiquitin (Ubi) puncta surround Hrs puncta; R–S” – BicD::GFP is not obviously punctate and fills the central space containing EPS15 puncta. Note the large central vesicle that excludes both markers in example R. T-T” – Rab5 puncta are found in the central space; U-U” – Rab6 is not obviously punctate and fills the central space. Several other Rab proteins do not enter the capes. Due to the widely varying extent and morphology of each cape, some panels show single confocal planes whereas others are maximum projections of up to 8 planes taken at 1 μm intervals. All scale bars represent 10 μm except for panels A”, T’, T” and U” which are 20 μm.