Figure S1.
MinSAT forward transcripts were depleted by RNA interference in MI oocytes. (A) MinSAT transcripts (forward and reverse) detected with strand-specific RNA FISH probes in MI oocytes (n = 8–12 oocytes combined from three independent experiments for each group). RNase treatment was performed as a negative control. Cen-ASO concentration was 40 µM. Scale bar: 5 µm. (B) The intensity of both forward and reverse transcript probes from A was compared with background subtraction. The integrated intensity (fluorescent foci) was measured specifically at the centromeric region (depicted by the yellow box in A). The number of fluorescent foci analyzed is indicated. Data were normalized with respect to the average intensity of forward transcripts (****, P < 0.0001, ANOVA with Tukey’s post hoc test). (C) Levels of MinSAT RNAs in GV, MI, and MII oocytes were tested by qRT-PCR (three independent experiments). Error bars are 95% confidence limits. 30 oocytes are in each group. (D) The knockdown efficiency of MajSAT-ASO was tested by qRT-PCR (ASO: 40 µM). All qRT-PCR tests were performed in three independent experiments. P values from 95% CI (*, P < 0.01). 30 oocytes in each group. (E) The integrity of centromeres shown in live oocytes. (F) MI completion rates were compared between control ASO– and MajSAT-ASO–injected oocytes (no significance, χ2 test). (G) MI completion rates were compared between control and α-amanitin (60 µM) treatment (no significance, χ2 test). (H) The knockdown efficiency of Cen-siRNA (40 µM) was tested by qRT-PCR. All qRT-PCR tests were performed in three independent experiments. P value and error bars from 95% CI (*, P < 0.01). 30 oocytes in each group. (I) Cen-RNA level was tested after siRNA interference by RNA FISH as in A. Scale bar: 5 µm. The fluorescent foci for measurements are indicated (yellow box; n = 6 oocytes combined from two independent experiments). (J) The intensity of Cen-RNA levels was measured as in B (****, P < 0.0001, t test). (K) MI completion rates in control siRNA– and Cen-siRNA–injected oocytes (**, P < 0.01, χ2 test). (L) MajSAT signal in an oocyte injected with control siRNA or Cen-siRNA. A damaged centromere is shown by the yellow box. (M) The incidence of damaged centromeres in oocytes injected with control siRNA or Cen-siRNA (****, P < 0.0001, t test). The average and standard error were calculated based on three independent experiments. Error bars indicate SD.

MinSAT forward transcripts were depleted by RNA interference in MI oocytes. (A) MinSAT transcripts (forward and reverse) detected with strand-specific RNA FISH probes in MI oocytes (n = 8–12 oocytes combined from three independent experiments for each group). RNase treatment was performed as a negative control. Cen-ASO concentration was 40 µM. Scale bar: 5 µm. (B) The intensity of both forward and reverse transcript probes from A was compared with background subtraction. The integrated intensity (fluorescent foci) was measured specifically at the centromeric region (depicted by the yellow box in A). The number of fluorescent foci analyzed is indicated. Data were normalized with respect to the average intensity of forward transcripts (****, P < 0.0001, ANOVA with Tukey’s post hoc test). (C) Levels of MinSAT RNAs in GV, MI, and MII oocytes were tested by qRT-PCR (three independent experiments). Error bars are 95% confidence limits. 30 oocytes are in each group. (D) The knockdown efficiency of MajSAT-ASO was tested by qRT-PCR (ASO: 40 µM). All qRT-PCR tests were performed in three independent experiments. P values from 95% CI (*, P < 0.01). 30 oocytes in each group. (E) The integrity of centromeres shown in live oocytes. (F) MI completion rates were compared between control ASO– and MajSAT-ASO–injected oocytes (no significance, χ2 test). (G) MI completion rates were compared between control and α-amanitin (60 µM) treatment (no significance, χ2 test). (H) The knockdown efficiency of Cen-siRNA (40 µM) was tested by qRT-PCR. All qRT-PCR tests were performed in three independent experiments. P value and error bars from 95% CI (*, P < 0.01). 30 oocytes in each group. (I) Cen-RNA level was tested after siRNA interference by RNA FISH as in A. Scale bar: 5 µm. The fluorescent foci for measurements are indicated (yellow box; n = 6 oocytes combined from two independent experiments). (J) The intensity of Cen-RNA levels was measured as in B (****, P < 0.0001, t test). (K) MI completion rates in control siRNA– and Cen-siRNA–injected oocytes (**, P < 0.01, χ2 test). (L) MajSAT signal in an oocyte injected with control siRNA or Cen-siRNA. A damaged centromere is shown by the yellow box. (M) The incidence of damaged centromeres in oocytes injected with control siRNA or Cen-siRNA (****, P < 0.0001, t test). The average and standard error were calculated based on three independent experiments. Error bars indicate SD.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal