Activation Assay:Article Title: The Critical Role of Enhanced OXPHOS and Mitochondrial Hyperpolarization in Simulated Microgravity‐Induced Oocyte Maturation Arrest
Article Snippet: .. Parthenogenetic activation was induced by placing in vitro matured oocytes in Ca 2 +‐free CZB medium (modified BMOC 2 medium) supplemented with 5 mM SrCl 2 (10025 70–4; Sangon Biotech, China) 5 mg mL −1 cytochalasin B (CB) for 6 h. Activated oocytes were then cultured in KSOM medium, and the development of two‐cell embryos and blastocysts was assessed at 24 and 96 h, respectively. .. Oocytes were fixed in 4% paraformaldehyde for 30 min, followed by permeabilization in 0.5% Triton X‐100 for 2 h. Samples were blocked in PBS containing 1% BSA, 0.1% Tween‐20, and 0.01% Triton X‐100 for 1 h and then incubated overnight at 4°C with primary antibodies: human anti‐centromere (1:500, Antibodies Incorporated 15‐234‐0001), mouse anti‐tubulin (1:300, Sigma, F2168), rabbit anti‐tubulin (1:500, Abcam ab6046), mouse anti‐pericentrin (1:400, BD Biosciences 611814), rabbit anti‐HSPE1 (1:300, Sigma–Aldrich ZRB1145), rabbit anti‐HSPD1 (1:300, Sigma‐Aldrich SAB4501465) and mouse anti‐HSPD1 (1:300, Absea KC‐6176).
In Vitro:Article Title: The Critical Role of Enhanced OXPHOS and Mitochondrial Hyperpolarization in Simulated Microgravity‐Induced Oocyte Maturation Arrest
Article Snippet: .. Parthenogenetic activation was induced by placing in vitro matured oocytes in Ca 2 +‐free CZB medium (modified BMOC 2 medium) supplemented with 5 mM SrCl 2 (10025 70–4; Sangon Biotech, China) 5 mg mL −1 cytochalasin B (CB) for 6 h. Activated oocytes were then cultured in KSOM medium, and the development of two‐cell embryos and blastocysts was assessed at 24 and 96 h, respectively. .. Oocytes were fixed in 4% paraformaldehyde for 30 min, followed by permeabilization in 0.5% Triton X‐100 for 2 h. Samples were blocked in PBS containing 1% BSA, 0.1% Tween‐20, and 0.01% Triton X‐100 for 1 h and then incubated overnight at 4°C with primary antibodies: human anti‐centromere (1:500, Antibodies Incorporated 15‐234‐0001), mouse anti‐tubulin (1:300, Sigma, F2168), rabbit anti‐tubulin (1:500, Abcam ab6046), mouse anti‐pericentrin (1:400, BD Biosciences 611814), rabbit anti‐HSPE1 (1:300, Sigma–Aldrich ZRB1145), rabbit anti‐HSPD1 (1:300, Sigma‐Aldrich SAB4501465) and mouse anti‐HSPD1 (1:300, Absea KC‐6176).
Modification:Article Title: The Critical Role of Enhanced OXPHOS and Mitochondrial Hyperpolarization in Simulated Microgravity‐Induced Oocyte Maturation Arrest
Article Snippet: .. Parthenogenetic activation was induced by placing in vitro matured oocytes in Ca 2 +‐free CZB medium (modified BMOC 2 medium) supplemented with 5 mM SrCl 2 (10025 70–4; Sangon Biotech, China) 5 mg mL −1 cytochalasin B (CB) for 6 h. Activated oocytes were then cultured in KSOM medium, and the development of two‐cell embryos and blastocysts was assessed at 24 and 96 h, respectively. .. Oocytes were fixed in 4% paraformaldehyde for 30 min, followed by permeabilization in 0.5% Triton X‐100 for 2 h. Samples were blocked in PBS containing 1% BSA, 0.1% Tween‐20, and 0.01% Triton X‐100 for 1 h and then incubated overnight at 4°C with primary antibodies: human anti‐centromere (1:500, Antibodies Incorporated 15‐234‐0001), mouse anti‐tubulin (1:300, Sigma, F2168), rabbit anti‐tubulin (1:500, Abcam ab6046), mouse anti‐pericentrin (1:400, BD Biosciences 611814), rabbit anti‐HSPE1 (1:300, Sigma–Aldrich ZRB1145), rabbit anti‐HSPD1 (1:300, Sigma‐Aldrich SAB4501465) and mouse anti‐HSPD1 (1:300, Absea KC‐6176).
Cell Culture:Article Title: The Critical Role of Enhanced OXPHOS and Mitochondrial Hyperpolarization in Simulated Microgravity‐Induced Oocyte Maturation Arrest
Article Snippet: .. Parthenogenetic activation was induced by placing in vitro matured oocytes in Ca 2 +‐free CZB medium (modified BMOC 2 medium) supplemented with 5 mM SrCl 2 (10025 70–4; Sangon Biotech, China) 5 mg mL −1 cytochalasin B (CB) for 6 h. Activated oocytes were then cultured in KSOM medium, and the development of two‐cell embryos and blastocysts was assessed at 24 and 96 h, respectively. .. Oocytes were fixed in 4% paraformaldehyde for 30 min, followed by permeabilization in 0.5% Triton X‐100 for 2 h. Samples were blocked in PBS containing 1% BSA, 0.1% Tween‐20, and 0.01% Triton X‐100 for 1 h and then incubated overnight at 4°C with primary antibodies: human anti‐centromere (1:500, Antibodies Incorporated 15‐234‐0001), mouse anti‐tubulin (1:300, Sigma, F2168), rabbit anti‐tubulin (1:500, Abcam ab6046), mouse anti‐pericentrin (1:400, BD Biosciences 611814), rabbit anti‐HSPE1 (1:300, Sigma–Aldrich ZRB1145), rabbit anti‐HSPD1 (1:300, Sigma‐Aldrich SAB4501465) and mouse anti‐HSPD1 (1:300, Absea KC‐6176).
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