Incubation:Article Title: Regulation of autophagy and its role in late preimplantation during mouse embryo development
Article Snippet: .. After washing in PBS-PVA, embryos were subsequently incubated with Alexa Fluor 568-labelled goat anti-mouse IgG secondary antibody (Invitrogen/Molecular Probes, Eugene, OR) at a 1:500 dilution for 2 h at room temperature. .. Finally, embryos were mounted on glass slides in Vecta-shield (Vector Laboratories Inc., Burlingame, CA) supplemented with 1 μg/ml 4’,6-diamidino-2-phenylindole (DAPI).
Article Title: Galangin Regulates Oxidative Stress Levels in Porcine Embryos Through Interaction with the Neh1 Domain of Nrf2
Article Snippet: .. The embryos were washed 3 times with PBS-PVA (0.1% PVA) and then incubated with 10 mM 2’,7’-dichlorodihydrofluorescein diacetate (Invitrogen, C400, Carlsbad, CA, USA) or 10 mM 4-chloromethyl-6,8-difluoro-7-hydroxycoumarin (Invitrogen, C12881 , Carlsbad, CA, USA) for 30 min. Photographs were taken using fluorescence microscopy (green fluorescence, 490 nm; blue fluorescence, 370 nm). .. Fluorescence intensity analysis was performed using ImageJ (v1.54e, National Institutes of Health, Bethesda, MD, USA).
Article Title: Melatonin counteracts Iodoacetic acid induced ferroptosis by regulating Nrf2 in porcine oocytes.
Article Snippet: .. After washing three times with PBS/PVA, oocytes were incubated at room temperature for 1 h with Alexa Fluor 488TM donkey anti-mouse IgG (H + L) (Invitrogen, A21202) or Alexa Fluor 546TM donkey anti-rabbit IgG (H + L) (Invitrogen, A10040 n). .. Subsequently, the oocytes were stained with 10 μg/mL DAPI for 10 min, washed three times with PBS/PVA, mounted onto slides, and examined under a confocal microscope (Zeiss LSM 710 META).
Article Title: Galangin Regulates Oxidative Stress Levels in Porcine Embryos Through Interaction with the Neh1 Domain of Nrf2.
Article Snippet: .. The embryos were washed 3 times with PBS-PVA (0.1% PVA) and then incubated with 10 mM 2’,7’-dichlorodihydrofluorescein diacetate (Invitrogen, C400, Carlsbad, CA, USA) or 10 mM 4-chloromethyl-6,8-difluoro-7-hydroxycoumarin (Invitrogen, C12881, Carlsbad, CA, USA) for 30 min. Photographs were taken using fluorescence microscopy (green fluorescence, 490 nm; blue fluorescence, 370 nm). .. Fluorescence intensity analysis was performed using ImageJ (v1.54e, National Institutes of Health, Bethesda, MD, USA).
Injection:Article Title: Quercetin ameliorates oxidative stress in BMP15-deficient oocytes by regulating the ERK1/2-GSK3β-CypD pathway.
Article Snippet: The primers used for PCR were synthesized by Sangon Biotech (Shanghai, China; Supplementary Table S3). .. The oocytes and CGCs were collected from the NC injection, ctRNP injection, and ctRNP injection supplementation of QUE groups, washed three times with PBS-PVA, and transferred into EP-tubes with 8 μL of cells-to-cDNATM II Kit (AM1722, Invitrogen, CA, USA). .. RNA was isolated and reverse-transcribed to cDNA using the SuperScriptTM II Reverse Transcriptase Kit (11750150; Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions.
Article Title: Quercetin ameliorates oxidative stress in BMP15 -deficient oocytes by regulating the ERK1/2-GSK3β-CypD pathway
Article Snippet: The primers used for PCR were synthesized by Sangon Biotech (Shanghai, China; ). .. The oocytes and CGCs were collected from the NC injection, ctRNP injection, and ctRNP injection supplementation of QUE groups, washed three times with PBS-PVA, and transferred into EP-tubes with 8 μL of cells-to-cDNATM II Kit (AM1722, Invitrogen, CA, USA). .. RNA was isolated and reverse-transcribed to cDNA using the SuperScriptTM II Reverse Transcriptase Kit (11750150; Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions.
Fluorescence:Article Title: Galangin Regulates Oxidative Stress Levels in Porcine Embryos Through Interaction with the Neh1 Domain of Nrf2
Article Snippet: .. The embryos were washed 3 times with PBS-PVA (0.1% PVA) and then incubated with 10 mM 2’,7’-dichlorodihydrofluorescein diacetate (Invitrogen, C400, Carlsbad, CA, USA) or 10 mM 4-chloromethyl-6,8-difluoro-7-hydroxycoumarin (Invitrogen, C12881 , Carlsbad, CA, USA) for 30 min. Photographs were taken using fluorescence microscopy (green fluorescence, 490 nm; blue fluorescence, 370 nm). .. Fluorescence intensity analysis was performed using ImageJ (v1.54e, National Institutes of Health, Bethesda, MD, USA).
Article Title: Galangin Regulates Oxidative Stress Levels in Porcine Embryos Through Interaction with the Neh1 Domain of Nrf2.
Article Snippet: .. The embryos were washed 3 times with PBS-PVA (0.1% PVA) and then incubated with 10 mM 2’,7’-dichlorodihydrofluorescein diacetate (Invitrogen, C400, Carlsbad, CA, USA) or 10 mM 4-chloromethyl-6,8-difluoro-7-hydroxycoumarin (Invitrogen, C12881, Carlsbad, CA, USA) for 30 min. Photographs were taken using fluorescence microscopy (green fluorescence, 490 nm; blue fluorescence, 370 nm). .. Fluorescence intensity analysis was performed using ImageJ (v1.54e, National Institutes of Health, Bethesda, MD, USA).
Microscopy:Article Title: Galangin Regulates Oxidative Stress Levels in Porcine Embryos Through Interaction with the Neh1 Domain of Nrf2
Article Snippet: .. The embryos were washed 3 times with PBS-PVA (0.1% PVA) and then incubated with 10 mM 2’,7’-dichlorodihydrofluorescein diacetate (Invitrogen, C400, Carlsbad, CA, USA) or 10 mM 4-chloromethyl-6,8-difluoro-7-hydroxycoumarin (Invitrogen, C12881 , Carlsbad, CA, USA) for 30 min. Photographs were taken using fluorescence microscopy (green fluorescence, 490 nm; blue fluorescence, 370 nm). .. Fluorescence intensity analysis was performed using ImageJ (v1.54e, National Institutes of Health, Bethesda, MD, USA).
Article Title: Galangin Regulates Oxidative Stress Levels in Porcine Embryos Through Interaction with the Neh1 Domain of Nrf2.
Article Snippet: .. The embryos were washed 3 times with PBS-PVA (0.1% PVA) and then incubated with 10 mM 2’,7’-dichlorodihydrofluorescein diacetate (Invitrogen, C400, Carlsbad, CA, USA) or 10 mM 4-chloromethyl-6,8-difluoro-7-hydroxycoumarin (Invitrogen, C12881, Carlsbad, CA, USA) for 30 min. Photographs were taken using fluorescence microscopy (green fluorescence, 490 nm; blue fluorescence, 370 nm). .. Fluorescence intensity analysis was performed using ImageJ (v1.54e, National Institutes of Health, Bethesda, MD, USA).
Staining:Article Title: Effects of L-Carnitine on the Developmental Competence of Bovine Oocytes.
Article Snippet: .. Then, these oocytes were washed 3 times in PBS-PVA 0.1% and stained with 10 μg/mL Nile red (Invitrogen, Carlsbad, CA, USA) for 4 h at room temperature. .. Subsequently, oocytes were washed 3 times in PBS-PVA 0.1%, mounted on glass slides, and examined by confocal microscopy (Leica, SP8, 63×, Buffalo Grove, IL, USA).
Article Title: Effects of L-Carnitine on the Developmental Competence of Bovine Oocytes
Article Snippet: .. Then, these oocytes were washed 3 times in PBS-PVA 0.1% and stained with 10 μg/mL Nile red (Invitrogen, Carlsbad, CA, USA) for 4 h at room temperature. .. Subsequently, oocytes were washed 3 times in PBS-PVA 0.1%, mounted on glass slides, and examined by confocal microscopy (Leica, SP8, 63×, Buffalo Grove, IL, USA).
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