Figure 6. The midbody is a site of spatiotemporally regulated translation which also occurs in different cell types.

(A) Translational onset (α-Puro; arrowheads at MB) occurred precisely as cells formally exited mitosis at the G1 transition, coincident with the mature reformation of the nuclear envelope (detected by lamin A/C) and the de-condensation of chromatin (DNA detected by DAPI staining). DAPI: 4’,6-diamidino-2-phenylindole. Quantification is noted at each stage in figure; 83% Early Telophase (n =10/12), 100% Late Telophase (n =3/3) and 100% MBR (n =3/3).

(B) Quantification of the number of distinct puromycin rings observed at different points during the late stages of mitosis, namely early telophase (ET), late telophase (LT), and MBR. The α-Puro label was primarily found in late telophase/G1 and continued in the MBR stage after MBR release. Quantification is noted next to each stage. Line scans denoted by the dotted line in each schematic was quantified for data sets and plotted (n=10). Here, the puromycin ring is seen prominently during Late Telophase and MBR stages. Asterix denotes significance.

(C) Retinal pigment epithelium cells (RPE)(n=5), HeLa CCL2 cells(n=5), CHO cells (n=4), and neural stem/progenitor cells (NSPCs)(n=4) all had puromycin rings labeled with MKLP1 within the bridge (tubulin). DAPI: 4’,6-diamidino-2-phenylindole. Quantification is noted next to each cell type, which are 100% for each cell type.

Scale bars are 1 μm unless noted.