Midbodies contain molecular machinery required for protein translation. (A) GFP-MKLP1 expressing HeLa cells were either mock-treated or treated with CHMP4B siRNA for 72 h and subjected to staining with smFISH probes against CHMP4B. Arrows in the inset point to CHMP4B mRNA particles. The cell outline is marked in yellow with the asterisks marking the MB. The black dashed square represents the region of the image used for the inset. Scale bars: 10 μm. (B) ImageJ was used to count the number of CHMP4B smFISH particles in the mock or CHMP4B siRNA-treated cells. Each dot represents a single cell. The means and SD were calculated from five randomly chosen cells in telophase. (C) The Database for Annotation, Visualization, and Integrated Discovery (DAVID) was used to determine whether ribosomal proteins and translation initiation factors are enriched in all three published MB proteomes. (D) HeLa cells were fixed and subjected to immunostaining with anti-acetylated-tubulin (red) and anti-RPL3 (green) antibodies. The line used for the line intensity graphs is marked in yellow and the asterisks mark the MB. The white dashed square represents the region of the image used for the inset. Scale bars: 10 μm. Inset scale bars: 2 μm. (E and F) Line intensity graphs representing the intensity of acetylated-tubulin (red) and RPL3 (green). Negative control was derived from samples that were treated with secondary antibodies but without anti-RPL3 antibodies.