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. 2022 Nov 15;6(1):e202201540. doi: 10.26508/lsa.202201540

Figure S5. Microinjection with a dimerization mutant protein cytATL R192Q.

Figure S5.

(A) Experimental layout to induce a metaphase arrest by microinjection of UbcH10C114S. After arrest (∼5 min), embryos were subjected to a second microinjection either with buffer or with cytATL R192Q protein. ER and spindle morphology were monitored by ERsp-EYFP–KDEL (green) and Alexa647-labelled tubulin (magenta), respectively. (B, C) Still images after microinjection of buffer (B) or the dimerization mutant protein cytATL R192Q (C) in metaphase-arrested embryos; time (min:s) is relative to the second microinjection (buffer/cytATL R192Q). Insets show a single nucleus. Scale bar is 10 μm. (D) ER exclusion area for control (+buffer) or dimerization mutant cytATL (+cytATL R192Q) conditions at the first (t = 0 min) and last (t = 10 min) time points of the time-lapse. (E, F) Quantitative analysis of spindle morphology by measuring spindle length (E) and spindle width (F) for control (+buffer) or dimerization mutant cytATL (+cytATL R192Q) conditions at the first (t = 0 min) and last (t = 10 min) time points. Statistical analysis using N = 3 embryos (buffer), N = 8 embryos (cytATL R192Q), n = 5 nuclei per embryo. Asterisks represent statistical significance derived from Kruskal–Wallis (Dunn’s multiple comparisons test) (D) or unpaired t test (two-sided). *P < 0.05, n.s., nonsignificant, P > 0.05.