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Aluminum exposure induces G2/M phase arrest in mouse zygotes (A) Representative images of γ-H2A.X in zygotes from the control group and the AlCl 3 -treated group by immunofluorescence staining. Green, γ-H2A.X; blue, DNA. Scale bars, 20 μm. (B) Relative fluorescence intensity of γ-H2A.X. Control, n = 40; AlCl 3 , n = 41; ∗∗ p < 0.005. (C) Expression level and statistical analysis of RAD51 protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗∗∗ p < 0.0001. (D) The mRNA expression levels of DNA damage-related genes were significantly downregulated in the treatment group compared with the control by RT-qPCR. ∗∗∗ p < 0.0005, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001. (E) Representative fluorescence images of CHK1 signals in zygotes of the control group and AlCl 3 -treated group by immunofluorescence staining. Green, CHK1; blue, DNA. Scale bars, 20 μm. (F) Relative fluorescence intensity of CHK1. Control, n = 34; AlCl 3 , n = 33; ∗∗ p < 0.005. (G) Expression level and statistical analysis of CDC25C protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗∗ p < 0.001. (H) Expression level and statistical analysis of Cyclin B1 protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗ p < 0.01. (I) Expression level and statistical analysis of p <t>-CDK1(T161)</t> protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗ p < 0.005. (J) The mRNA expression of cell cycle regulatory genes by RT-qPCR. ∗∗∗ p < 0.001, ∗∗ p < 0.005, ∗∗ p < 0.005, ∗ p < 0.05. Data are presented as mean ± SEM. All experiments were performed in three independent biological replicates, with three technical replicates per biological replicate; mouse zygotes were used as research subjects. n represents the number of zygotes analyzed. Statistical analysis was performed using paired t tests. Asterisks indicate statistical significance: ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.
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Aluminum exposure induces G2/M phase arrest in mouse zygotes (A) Representative images of γ-H2A.X in zygotes from the control group and the AlCl 3 -treated group by immunofluorescence staining. Green, γ-H2A.X; blue, DNA. Scale bars, 20 μm. (B) Relative fluorescence intensity of γ-H2A.X. Control, n = 40; AlCl 3 , n = 41; ∗∗ p < 0.005. (C) Expression level and statistical analysis of RAD51 protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗∗∗ p < 0.0001. (D) The mRNA expression levels of DNA damage-related genes were significantly downregulated in the treatment group compared with the control by RT-qPCR. ∗∗∗ p < 0.0005, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001. (E) Representative fluorescence images of CHK1 signals in zygotes of the control group and AlCl 3 -treated group by immunofluorescence staining. Green, CHK1; blue, DNA. Scale bars, 20 μm. (F) Relative fluorescence intensity of CHK1. Control, n = 34; AlCl 3 , n = 33; ∗∗ p < 0.005. (G) Expression level and statistical analysis of CDC25C protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗∗ p < 0.001. (H) Expression level and statistical analysis of Cyclin B1 protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗ p < 0.01. (I) Expression level and statistical analysis of p <t>-CDK1(T161)</t> protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗ p < 0.005. (J) The mRNA expression of cell cycle regulatory genes by RT-qPCR. ∗∗∗ p < 0.001, ∗∗ p < 0.005, ∗∗ p < 0.005, ∗ p < 0.05. Data are presented as mean ± SEM. All experiments were performed in three independent biological replicates, with three technical replicates per biological replicate; mouse zygotes were used as research subjects. n represents the number of zygotes analyzed. Statistical analysis was performed using paired t tests. Asterisks indicate statistical significance: ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.
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Aluminum exposure induces G2/M phase arrest in mouse zygotes (A) Representative images of γ-H2A.X in zygotes from the control group and the AlCl 3 -treated group by immunofluorescence staining. Green, γ-H2A.X; blue, DNA. Scale bars, 20 μm. (B) Relative fluorescence intensity of γ-H2A.X. Control, n = 40; AlCl 3 , n = 41; ∗∗ p < 0.005. (C) Expression level and statistical analysis of RAD51 protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗∗∗ p < 0.0001. (D) The mRNA expression levels of DNA damage-related genes were significantly downregulated in the treatment group compared with the control by RT-qPCR. ∗∗∗ p < 0.0005, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001. (E) Representative fluorescence images of CHK1 signals in zygotes of the control group and AlCl 3 -treated group by immunofluorescence staining. Green, CHK1; blue, DNA. Scale bars, 20 μm. (F) Relative fluorescence intensity of CHK1. Control, n = 34; AlCl 3 , n = 33; ∗∗ p < 0.005. (G) Expression level and statistical analysis of CDC25C protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗∗ p < 0.001. (H) Expression level and statistical analysis of Cyclin B1 protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗ p < 0.01. (I) Expression level and statistical analysis of p <t>-CDK1(T161)</t> protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗ p < 0.005. (J) The mRNA expression of cell cycle regulatory genes by RT-qPCR. ∗∗∗ p < 0.001, ∗∗ p < 0.005, ∗∗ p < 0.005, ∗ p < 0.05. Data are presented as mean ± SEM. All experiments were performed in three independent biological replicates, with three technical replicates per biological replicate; mouse zygotes were used as research subjects. n represents the number of zygotes analyzed. Statistical analysis was performed using paired t tests. Asterisks indicate statistical significance: ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.
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Aluminum exposure induces G2/M phase arrest in mouse zygotes (A) Representative images of γ-H2A.X in zygotes from the control group and the AlCl 3 -treated group by immunofluorescence staining. Green, γ-H2A.X; blue, DNA. Scale bars, 20 μm. (B) Relative fluorescence intensity of γ-H2A.X. Control, n = 40; AlCl 3 , n = 41; ∗∗ p < 0.005. (C) Expression level and statistical analysis of RAD51 protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗∗∗ p < 0.0001. (D) The mRNA expression levels of DNA damage-related genes were significantly downregulated in the treatment group compared with the control by RT-qPCR. ∗∗∗ p < 0.0005, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001. (E) Representative fluorescence images of CHK1 signals in zygotes of the control group and AlCl 3 -treated group by immunofluorescence staining. Green, CHK1; blue, DNA. Scale bars, 20 μm. (F) Relative fluorescence intensity of CHK1. Control, n = 34; AlCl 3 , n = 33; ∗∗ p < 0.005. (G) Expression level and statistical analysis of CDC25C protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗∗ p < 0.001. (H) Expression level and statistical analysis of Cyclin B1 protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗ p < 0.01. (I) Expression level and statistical analysis of p <t>-CDK1(T161)</t> protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗ p < 0.005. (J) The mRNA expression of cell cycle regulatory genes by RT-qPCR. ∗∗∗ p < 0.001, ∗∗ p < 0.005, ∗∗ p < 0.005, ∗ p < 0.05. Data are presented as mean ± SEM. All experiments were performed in three independent biological replicates, with three technical replicates per biological replicate; mouse zygotes were used as research subjects. n represents the number of zygotes analyzed. Statistical analysis was performed using paired t tests. Asterisks indicate statistical significance: ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.
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Aluminum exposure induces G2/M phase arrest in mouse zygotes (A) Representative images of γ-H2A.X in zygotes from the control group and the AlCl 3 -treated group by immunofluorescence staining. Green, γ-H2A.X; blue, DNA. Scale bars, 20 μm. (B) Relative fluorescence intensity of γ-H2A.X. Control, n = 40; AlCl 3 , n = 41; ∗∗ p < 0.005. (C) Expression level and statistical analysis of RAD51 protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗∗∗ p < 0.0001. (D) The mRNA expression levels of DNA damage-related genes were significantly downregulated in the treatment group compared with the control by RT-qPCR. ∗∗∗ p < 0.0005, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001. (E) Representative fluorescence images of CHK1 signals in zygotes of the control group and AlCl 3 -treated group by immunofluorescence staining. Green, CHK1; blue, DNA. Scale bars, 20 μm. (F) Relative fluorescence intensity of CHK1. Control, n = 34; AlCl 3 , n = 33; ∗∗ p < 0.005. (G) Expression level and statistical analysis of CDC25C protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗∗ p < 0.001. (H) Expression level and statistical analysis of Cyclin B1 protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗ p < 0.01. (I) Expression level and statistical analysis of p <t>-CDK1(T161)</t> protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗ p < 0.005. (J) The mRNA expression of cell cycle regulatory genes by RT-qPCR. ∗∗∗ p < 0.001, ∗∗ p < 0.005, ∗∗ p < 0.005, ∗ p < 0.05. Data are presented as mean ± SEM. All experiments were performed in three independent biological replicates, with three technical replicates per biological replicate; mouse zygotes were used as research subjects. n represents the number of zygotes analyzed. Statistical analysis was performed using paired t tests. Asterisks indicate statistical significance: ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.
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Aluminum exposure induces G2/M phase arrest in mouse zygotes (A) Representative images of γ-H2A.X in zygotes from the control group and the AlCl 3 -treated group by immunofluorescence staining. Green, γ-H2A.X; blue, DNA. Scale bars, 20 μm. (B) Relative fluorescence intensity of γ-H2A.X. Control, n = 40; AlCl 3 , n = 41; ∗∗ p < 0.005. (C) Expression level and statistical analysis of RAD51 protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗∗∗ p < 0.0001. (D) The mRNA expression levels of DNA damage-related genes were significantly downregulated in the treatment group compared with the control by RT-qPCR. ∗∗∗ p < 0.0005, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001. (E) Representative fluorescence images of CHK1 signals in zygotes of the control group and AlCl 3 -treated group by immunofluorescence staining. Green, CHK1; blue, DNA. Scale bars, 20 μm. (F) Relative fluorescence intensity of CHK1. Control, n = 34; AlCl 3 , n = 33; ∗∗ p < 0.005. (G) Expression level and statistical analysis of CDC25C protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗∗ p < 0.001. (H) Expression level and statistical analysis of Cyclin B1 protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗ p < 0.01. (I) Expression level and statistical analysis of p -CDK1(T161) protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗ p < 0.005. (J) The mRNA expression of cell cycle regulatory genes by RT-qPCR. ∗∗∗ p < 0.001, ∗∗ p < 0.005, ∗∗ p < 0.005, ∗ p < 0.05. Data are presented as mean ± SEM. All experiments were performed in three independent biological replicates, with three technical replicates per biological replicate; mouse zygotes were used as research subjects. n represents the number of zygotes analyzed. Statistical analysis was performed using paired t tests. Asterisks indicate statistical significance: ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Journal: iScience

Article Title: Aluminum exposure impairs nuclear envelope breakdown for mouse zygote formation

doi: 10.1016/j.isci.2026.115807

Figure Lengend Snippet: Aluminum exposure induces G2/M phase arrest in mouse zygotes (A) Representative images of γ-H2A.X in zygotes from the control group and the AlCl 3 -treated group by immunofluorescence staining. Green, γ-H2A.X; blue, DNA. Scale bars, 20 μm. (B) Relative fluorescence intensity of γ-H2A.X. Control, n = 40; AlCl 3 , n = 41; ∗∗ p < 0.005. (C) Expression level and statistical analysis of RAD51 protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗∗∗ p < 0.0001. (D) The mRNA expression levels of DNA damage-related genes were significantly downregulated in the treatment group compared with the control by RT-qPCR. ∗∗∗ p < 0.0005, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001. (E) Representative fluorescence images of CHK1 signals in zygotes of the control group and AlCl 3 -treated group by immunofluorescence staining. Green, CHK1; blue, DNA. Scale bars, 20 μm. (F) Relative fluorescence intensity of CHK1. Control, n = 34; AlCl 3 , n = 33; ∗∗ p < 0.005. (G) Expression level and statistical analysis of CDC25C protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗∗ p < 0.001. (H) Expression level and statistical analysis of Cyclin B1 protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗ p < 0.01. (I) Expression level and statistical analysis of p -CDK1(T161) protein in zygotes of the control group and AlCl 3 -treated group by western blot. ∗∗ p < 0.005. (J) The mRNA expression of cell cycle regulatory genes by RT-qPCR. ∗∗∗ p < 0.001, ∗∗ p < 0.005, ∗∗ p < 0.005, ∗ p < 0.05. Data are presented as mean ± SEM. All experiments were performed in three independent biological replicates, with three technical replicates per biological replicate; mouse zygotes were used as research subjects. n represents the number of zygotes analyzed. Statistical analysis was performed using paired t tests. Asterisks indicate statistical significance: ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Anti-phospho-CDK1 (Thr161) antibody , HuaBio , Cat# HA721987; RRID: N/A.

Techniques: Control, Immunofluorescence, Staining, Fluorescence, Expressing, Western Blot, Quantitative RT-PCR