Science - USA (2022-02-11)

fluorescence-minus-one (FMO) controls cor- respond to cells that were stained with mAbs for the detection of lineage markers but ...

Enumeration of cells via flow cytometry was performed using a defined number of Sphero blank calibration beads (BD BioSciences). ...

12% Bis-Tris Plus) and transferred to iBlot 2 PVDF membranes with an iBlot 2 system according to manufacturer’s instructions (al ...

Acad. Sci. U.S.A. 93 , 3357–3361 (1996). doi:10.1073/ pnas.93.8.3357; pmid: 8622941 D. Vremec, K. Shortman, The isolation and i ...

RESEARCH ARTICLE SUMMARY ◥ CELL BIOLOGY Mechanism of spindle pole organization and instability in human oocytes Chun So, Katerin ...

RESEARCH ARTICLE ◥ CELL BIOLOGY Mechanism of spindle pole organization and instability in human oocytes Chun So^1 , Katerina Men ...

to the organization of spindle poles in human oocytes. To test directly if NUMA is required for spindle pole organization, we de ...

no longer had NUMA at the spindle poles, and the poles became fully defocused (Fig. 1, G to I). Thus, NUMA is required for pole ...

reexpressed four different NUMA mutants in aMTOC-free mouse oocytes (fig. S6, A to C). NUMA(DN) and NUMA(SpM-4A), which cannot i ...

constitutive and acute depletion of LIS1 pheno- copied dynein-dynactin inhibition and led to spindle pole defocusing in aMTOC-fr ...

Soet al.,Science 375 , eabj3944 (2022) 11 February 2022 6 of 19 Microtubules Dynactin Prophase Nuclear envelope breakdown Onset ...

assembled multipolar spindles and spindles with broad poles, respectively (Fig. 5, F to H; and fig. S14, A and B). By contrast, ...

45% of KIFC1-depleted bovine oocytes under- went tridirectional division (Fig. 6C), which was likely because fewer oocytes resto ...

analysis revealed that wild-type and aMTOC- free mouse oocytes had a comparable fraction of mobile mClover3-KIFC1, but the turno ...

Unlike the other two kinesin-14 family mem- bers KIFC2 and KIFC3, KIFC1 distinctively possesses a microtubule-binding domain at ...

KIFC1’s functions in aMTOC-free spindles, we depleted endogenous KIFC1 and reexpressed wild-type KIFC1 and KIFC1(N593K), which c ...

Discussion Our data elucidate a cause of spindle instability in human oocytes: Human oocytes are deficient in the molecular moto ...

Preparation and culture of bovine and porcine oocytes All ovaries were obtained from local slaughter- houses. Bovine and porcine ...

the mRNA for 12 hours before release. For live imaging of the spindle assembly with Trim- Away, bovine oocytes were microinjecte ...

with 4000 U/ml lipase fromCandida rugose (Sigma-Aldrich) in 400 mM NaCl, 50 mM Tris (pH 7.2), 5 mM CaCl 2 , and 0.2% sodium taur ...