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. 2020 Mar 9;11:1269. doi: 10.1038/s41467-020-15002-8

Fig. 5. Optogenetic clustering of Rab11-associated vesicles results in failed abscission in vitro and in vivo.

Fig. 5

a, b Time-lapse of cytokinetic HeLa cells transfected with CRY2-mCherry and CIB1-mCerulean-Rab11 (black) in the absence (a) or presence of 488 nm light (b). Bar, 10 μm. Note the cleavage events of cytokinetic bridge (blue arrows, a), but not in b. c Bar graph depicting the percentage of total HeLa cells displaying a binucleate phenotype after being released from a metaphase synchronization for 2 h in the presence or absence of 488 nm light. Cells were transfected with CRY2-mCherry and CIB1-mCerulean-Rab11 as in a. Unpaired, two-tailed Mann–Whitney test, **p = 0.0043. Mean displayed ± SEM. n = 100 cells per treatment for n > 5 experiments. Dots represent individual values. Statistical results detailed in Supplementary Table 5. d A 3D rendering of embryos expressing CRY2 and CIB1-mCherry-Rab11 in the absence (left) and presence (right) of 488 nm light. Sox17:GFP-CAAX (magenta), CIB1-mCherry-Rab11 (cyan), and nuclei (DAPI—white) shown. Bar, 5 μm. e Representative images of single nuclei, binucleate, or multinucleate cells. Nuclei shown in grayscale (DAPI). Bar, 5 μm. f Bar graph depicting percentage of binucleate and/or multinucleate cells per KV in uninjected embryos and embryos expressing CIB1-mCherry-Rab11 or CRY2 and CIB1-mCherry-Rab11 plus or minus 488 nm light. One-way ANOVA with Dunnett’s multiple comparison test used, compared with uninjected embryos in the absence of 488 nm light exposure. Statistical results detailed in Supplementary Table 5. Analyses performed in n > 5 embryos over three experiments. Mean displayed ± SEM. Dots represent individual values.