Figure S5.

Effects of KASH5-DN, nocodazole, and P150-CC1 treatments in bovine zygotes, related to Figure 4

(A) Representative image of a bovine zygote expressing mClover3-KASH5-DN (green) and H2B-mScarlet (DNA, magenta).

(B-G) Representative images and quantification of zygotes in indicated groups displaying, upon KASH5-DN treatment, detached centrosomes (B and C), pronuclear migration defects (D and E) or separate spindles at anaphase onset (F and G). White, microtubules (mClover3-MAP4-MTBD). Magenta, DNA (H2B-mScarlet). Arrows indicate detached centrosomes and dashed lines indicate pronuclear envelopes. Z-projection, respectively 4, 7, 5, 8, 4, and 10 sections every 2.50 μm.

(H) Nuclear and surface occupancy indices in zygotes expressing GST or KASH5-DN.

(I) Zygotes treated with DMSO or nocodazole before pronuclear juxtaposition were scored for pronuclear migration defects.

(J) Nuclear and surface occupancy indices in zygotes treated with DMSO or nocodazole before pronuclear juxtaposition.

(K-L) Inner chromatin fraction and max chromatin polarity indices in zygotes treated with DMSO or nocodazole after pronuclear juxtaposition.

(M) Nuclear and surface occupancy indices in zygotes injected with BSA or P150-CC1.

Data are from four (C, E, G, H, M), six (I,J), three (K,L) independent experiments. The number of analyzed zygotes (C, E, G, I) and pronuclei (H, J, K, L, M) are specified in italics. p values were calculated using Fisher’s exact test (C, E, G, I) and unpaired two-tailed Student’s t test (H, J, K, L, M). Scale bars, 10 μm.