muc1 knockout mice  (Shanghai Model Organisms Center)

Journal: Cells

Article Title: MUC1 Protects Preimplantation Embryos In Vitro via Clearance of ROS by Triggering Mitophagy

doi: 10.3390/cells15090806

Figure Lengend Snippet: Expression and localization of MUC1 in gametes and preimplantation embryos. ( A – D ) Representative immunofluorescence images stained with antibody against MUC1 (green) and co-stained with DAPI (to stain DNA, blue) in human oocyte, mouse oocyte, human sperm, and mouse sperm, respectively. Magnified immunofluorescence image of human sperm is presented within the white box in ( C ). ( E ) Representative images stained with antibody against MUC1 (red) and co-stained with DAPI (to stain DNA, blue) from the 2PN stage to the blastocyst stage. NCtrl, negative control (no primary antibody). Scale bars: 20 µm. ( F ) Quantification of MUC1 fluorescence intensity using ImageJ. Number of samples analyzed at each stage: 2PN = 23, 2CELL = 25, 4CELL = 23, 8CELL = 23, MORULA = 19, and BLASTO = 29 (analyzed using ANOVA). ( G ) Real-time quantitative PCR results for Muc1 mRNA expression levels during early mouse embryonic development. Relative expression level of Muc1 is presented as fold change normalized to β-ACTIN. Number of samples analyzed at each stage: 2PN = 20, 2CELL = 16, 4CELL = 23, 8CELL = 17, MORULA = 17, and BLASTO = 15 (analyzed using ANOVA). Data in ( F , G ) are means ± SDs from at least three independent experiments.

Article Snippet: Wild-type and Muc1 knockout mice, both on the C57BL/6J background, were generated by the Shanghai Model Organisms Center, Inc. (Shanghai, China), as previously described [ ].

Techniques: Expressing, Immunofluorescence, Staining, Negative Control, Fluorescence, Real-time Polymerase Chain Reaction

Journal: Cells

Article Title: MUC1 Protects Preimplantation Embryos In Vitro via Clearance of ROS by Triggering Mitophagy

doi: 10.3390/cells15090806

Figure Lengend Snippet: Muc1 knockout has little, if any, effect on fertility in mice. ( A ) Genotyping of wild-type and Muc1 knockout mice. The black arrows indicate the target bands. ( B ) RT-qPCR analysis of wild-type and Muc1 knockout ovarian tissues. Relative expression level of Muc1 is presented as fold change normalized to β-ACTIN (analyzed using two-tailed Student’s t -test). ( C ) Representative immunofluorescence images depicting MUC1 expression in MII oocytes of wild-type and Muc1 knockout mice. MII, metaphase II. Scale bar: 20 µm. ( D ) Quantification of relative MUC1 fluorescence intensity in MII oocytes of wild-type and Muc1 knockout mice. Number of samples: wild-type = 33 and Muc1 knockout = 28 (analyzed using two-tailed Student’s t -test). ( E ) Litter size of wild-type and Muc1 knockout pairs (analyzed using two-tailed Student’s t -test). ( F ) Survival rate of pups of wild-type and Muc1 knockout pairs (analyzed using two-tailed Student’s t -test). ( G ) Offspring sex ratio from wild-type and Muc1 knockout pairs (analyzed using Chi-square test). ( H ) Interpregnancy interval of wild-type and Muc1 knockout pairs for same parents ( left ) and parturition ( right ) (analyzed using two-tailed Student’s t -test). Data in ( B , D ) are means ± SDs from at least three independent experiments. ns: not significant.

Article Snippet: Wild-type and Muc1 knockout mice, both on the C57BL/6J background, were generated by the Shanghai Model Organisms Center, Inc. (Shanghai, China), as previously described [ ].

Techniques: Knock-Out, Quantitative RT-PCR, Expressing, Two Tailed Test, Immunofluorescence, Fluorescence

Journal: Cells

Article Title: MUC1 Protects Preimplantation Embryos In Vitro via Clearance of ROS by Triggering Mitophagy

doi: 10.3390/cells15090806

Figure Lengend Snippet: Muc1 knockout impairs in vitro embryonic development in mice. ( A ) Overview of the experimental design (created in BioRender; https://BioRender.com/2uu2xma (accessed on 25th February 2026)). ( B ) Morphological comparison of D0.5, D1.5, D2.5, D3.0, D3.5, and D4.5 wild-type and Muc1 knockout embryos. Scale bar: 50 µm. White arrows indicate embryos with developmental arrest. ( C ) The rate of embryos that reached the 4-cell stage in wild-type and Muc1 knockout groups. Number of samples: wild-type = 80 and Muc1 knockout = 69 (analyzed using Chi-square test). ( D ) The rate of embryos that reached the blastocyst stage in wild-type and Muc1 knockout groups. Number of samples: wild-type = 80 and Muc1 knockout = 69 (analyzed using Chi-square test). ( E ) Representative immunofluorescence images stained with antibodies against Oct4 (green, ICM) and CDX2 (red, TE), and co-stained with DAPI (blue, nuclei). Scale bar: 10 µm. ( F ) The number of cells in blastocysts (ICM, TE, or total) of wild-type and Muc1 knockout groups based on three-dimensional reconstruction. Number of samples: wild-type = 20 and Muc1 knockout = 20 (analyzed using two-tailed Student’s t -test). ( G ) Representative immunofluorescence images stained with antibodies against Oct4 (green) and MUC1 (red), and co-stained with DAPI (blue). Scale bar: 10 µm. ( H ) Relative telomere length of wild-type and Muc1 knockout blastocysts. Number of samples: wild-type = 27 and Muc1 knockout = 27 (analyzed using two-tailed Student’s t -test). Data in ( C , D , F , H ) are means ± SDs from at least three independent experiments.

Article Snippet: Wild-type and Muc1 knockout mice, both on the C57BL/6J background, were generated by the Shanghai Model Organisms Center, Inc. (Shanghai, China), as previously described [ ].

Techniques: Knock-Out, In Vitro, Comparison, Immunofluorescence, Staining, Two Tailed Test

Journal: Cells

Article Title: MUC1 Protects Preimplantation Embryos In Vitro via Clearance of ROS by Triggering Mitophagy

doi: 10.3390/cells15090806

Figure Lengend Snippet: Muc1 knockout leads to accumulation of mtROS and damaged mitochondria. ( A ) Immunofluorescence staining of MitoSOX (red) and DNA (blue) ( left ), and statistical analysis of MitoSOX fluorescence intensity ( right ) in wild-type and Muc1 knockout groups at the blastocyst stage. Scale bar: 10 µm. Number of samples: wild-type = 22 and Muc1 knockout = 20 (analyzed using two-tailed Student’s t -test). ( B ) Immunofluorescence staining of MitoSOX (red) and DNA (blue) ( left ), and statistical analysis of MitoSOX fluorescence intensity ( right ) in wild-type and Muc1 knockout groups at the 4-cell stage. Scale bar: 10 µm. Number of samples: wild-type = 23 and Muc1 knockout = 20 (analyzed using two-tailed Student’s t -test). ( C ) Immunofluorescence staining of JC-1 (mitochondrial membrane potential) ( left ), and statistical analysis of JC-1 (red-to-green ratio) ( right ) in wild-type and Muc1 knockout groups at the blastocyst stage. Scale bar: 10 µm. Number of samples: wild-type = 23 and Muc1 knockout = 21 (analyzed using two-tailed Student’s t -test). ( D ) Immunofluorescence staining of p62 (green) and DNA (blue) ( left ), and statistical analysis of p62 fluorescence intensity ( right ) in wild-type and Muc1 knockout groups at the blastocyst stage. Scale bar: 10 µm. Number of samples: wild-type = 22 and Muc1 knockout = 21 (analyzed using two-tailed Student’s t -test). Data in ( A – D ) are means ± SDs from at least three independent experiments.

Article Snippet: Wild-type and Muc1 knockout mice, both on the C57BL/6J background, were generated by the Shanghai Model Organisms Center, Inc. (Shanghai, China), as previously described [ ].

Techniques: Knock-Out, Immunofluorescence, Staining, Fluorescence, Two Tailed Test, Membrane

Journal: Cells

Article Title: MUC1 Protects Preimplantation Embryos In Vitro via Clearance of ROS by Triggering Mitophagy

doi: 10.3390/cells15090806

Figure Lengend Snippet: Muc1 knockout leads to reduction in mitophagy. ( A ) Immunofluorescence staining of LC3 and MitoTracker in wild-type and Muc1 knockout groups at the blastocyst stage. Blue, DNA; green, LC3; red, MitoTracker. Scale bar: 10 µm. White arrows indicate the co-localization of LC3 and MitoTracker. ( B ) Pearson correlation coefficient for co-localization of LC3 and MitoTracker at the blastocyst stage. Number of samples: wild-type = 28, Muc1 knockout = 27 (analyzed using two-tailed Student’s t -test). ( C ) Pearson correlation coefficient for co-localization of LysoTracker and MitoTracker at the blastocyst stage. Number of samples: wild-type = 22 and Muc1 knockout = 21 (analyzed using two-tailed Student’s t -test). ( D ) Immunofluorescence staining of LysoTracker and MitoTracker in wild-type and Muc1 knockout groups at the blastocyst stage. Blue, DNA; green, MitoTracker; red, LysoTracker. Scale bar: 10 µm. White arrows indicate co-localization of LysoTracker and MitoTracker. ( E ) Immunofluorescence staining of LC3 and MitoTracker in wild-type and Muc1 knockout groups at the 4-cell stage. Blue, DNA; green, LC3; red, MitoTracker. Scale bar: 20 µm. White arrows indicate co-localization of LC3 and MitoTracker. ( F ) Pearson correlation coefficient for co-localization of LC3 and MitoTracker at the 4-cell stage. Number of samples: wild-type = 21, Muc1 knockout = 20 (analyzed using two-tailed Student’s t -test). ( G ) Immunofluorescence staining of PINK1 (red) and DNA (blue) ( left ), and statistical analysis of PINK1 fluorescence intensity ( right ) in wild-type and Muc1 knockout groups at the blastocyst stage. Scale bar: 10 µm. Number of samples: wild-type = 21 and Muc1 knockout = 20 (analyzed using two-tailed Student’s t -test). ( H ) Immunofluorescence staining of Parkin (red) and DNA (blue) ( left ), and statistical analysis of Parkin fluorescence intensity ( right ) in wild-type and Muc1 knockout groups at the blastocyst stage. Scale bar: 10 µm. Number of samples: wild-type = 20 and Muc1 knockout = 20 (analyzed using two-tailed Student’s t -test). Data in ( B , C , F – H ) are means ± SDs from at least three independent experiments.

Article Snippet: Wild-type and Muc1 knockout mice, both on the C57BL/6J background, were generated by the Shanghai Model Organisms Center, Inc. (Shanghai, China), as previously described [ ].

Techniques: Knock-Out, Immunofluorescence, Staining, Two Tailed Test, Fluorescence

Journal: Cells

Article Title: MUC1 Protects Preimplantation Embryos In Vitro via Clearance of ROS by Triggering Mitophagy

doi: 10.3390/cells15090806

Figure Lengend Snippet: Low-dose CCCP treatment rescues impaired mitophagy and blastocyst formation defects caused by Muc1 knockout. ( A ) Morphological comparison of D0.5, D1.5, D3.0, D3.5, and D4.5 WT, KO, WT + CCCP, and KO + CCCP embryos. Scale bar: 50 µm. White arrows indicate embryos with developmental arrest. ( B ) Blastocyst formation rates in WT, KO, WT + CCCP, and KO + CCCP groups. Number of samples analyzed in each group: WT = 37, KO = 45, WT + CCCP = 37, and KO + CCCP = 48 (analyzed using Chi-square test). ( C ) Immunofluorescence staining of MitoSOX in WT, KO, WT + CCCP, and KO + CCCP groups at the blastocyst stage. Blue, DNA; red, MitoSOX. Scale bar: 10 µm. ( D ) Relative fluorescence intensity of MitoSOX at the blastocyst stage. Number of samples analyzed in each group was as follows: WT = 20, KO = 21, WT + CCCP = 20, and KO + CCCP = 20 (analyzed using ANOVA). ( E ) Immunofluorescence staining of LC3 and MitoTracker in WT, KO, WT + CCCP, and KO + CCCP groups at the blastocyst stage. Blue, DNA; green, LC3; red, MitoTracker. Scale bar: 10 µm. White arrows indicate the co-localization of LC3 and MitoTracker. ( F ) Pearson correlation coefficient for co-localization of LC3 and MitoTracker at the blastocyst stage. Number of samples analyzed in each group: WT = 23, KO = 20, WT + CCCP = 22, and KO + CCCP = 20 (analyzed using ANOVA). Data in ( B , D , F ) are means ± SDs from at least three independent experiments. ns: not significant.

Article Snippet: Wild-type and Muc1 knockout mice, both on the C57BL/6J background, were generated by the Shanghai Model Organisms Center, Inc. (Shanghai, China), as previously described [ ].

Techniques: Knock-Out, Comparison, Immunofluorescence, Staining, Fluorescence

Journal: Cells

Article Title: MUC1 Protects Preimplantation Embryos In Vitro via Clearance of ROS by Triggering Mitophagy

doi: 10.3390/cells15090806

Figure Lengend Snippet: Vitamin C treatment restores abnormal embryonic development induced by Muc1 knockout by normalizing mtROS levels. ( A ) Morphological comparison of D0.5, D1.5, and D4.5 WT, KO, WT + VC, and KO + VC embryos. Scale bar: 50 µm. White arrows indicate embryos with developmental arrest. ( B ) Blastocyst formation rates in WT, KO, WT + VC, and KO + VC groups. Number of samples analyzed in each group: WT = 35, KO = 35, WT + VC = 34, and KO + VC = 37 (analyzed using Chi-square test). ( C ) Immunofluorescence staining of MitoSOX in WT, KO, WT + VC, and KO + VC groups at the blastocyst stage. Blue, DNA; red, MitoSOX. Scale bar: 10 µm. ( D ) Relative fluorescence intensity of MitoSOX at the blastocyst stage. Number of samples: WT = 22, KO = 20, WT + VC = 20, and KO + VC = 20 (analyzed using ANOVA). ( E ) Schematic overview of the working model (created in BioRender. https://BioRender.com/z21zhau (accessed on 28th February 2026)). The dashed arrow indicates partial blockade of the process. Embryos cultured in vitro are unavoidably exposed to physical stressors, inducing the generation of mtROS. MUC1 maintains the normal progression of early embryonic development by promoting mitophagy, which suppresses mtROS accumulation. Muc1 depletion impairs trophectoderm development and shortens telomere length. Low-dose CCCP and vitamin C treatment restore embryonic development by enhancing mitophagy and clearing mtROS, respectively. Data in ( B , D ) are means ± SDs from at least three independent experiments. ns: not significant.

Article Snippet: Wild-type and Muc1 knockout mice, both on the C57BL/6J background, were generated by the Shanghai Model Organisms Center, Inc. (Shanghai, China), as previously described [ ].

Techniques: Knock-Out, Comparison, Immunofluorescence, Staining, Fluorescence, Cell Culture, In Vitro