Extended Data Fig. 6: The capacity of early mouse embryos to degrade CISD1-Binding-labelled mitochondria.
From: Reduction of mtDNA heteroplasmy in mitochondrial replacement therapy by inducing forced mitophagy
a. Mouse embryos were injected with or without (control) CISD1-Binding mRNA at the zygote stage, cultured to different stages, and then the whole embryos were subjected to absolute quantification of the mtDNA. Data were analyzed by two-tailed unpaired t-test and expressed as average ± SEM. Each point represents one embryo from three biological replicates. For Zygote, n = 14, the values are 2.44 ± 0.1618 (control) and 2.485 ± 0.1586 (CISD1-Binding). For 2-Cell, n = 13, the values are 2.543 ± 0.1016 (control) and 2.642 ± 0.1891 (CISD1-Binding). For 4-Cell, n = 12 (control) and n = 11 (CISD1-Binding), the values are 2.529 ± 0.07611 (control) and 2.405 ± 0.1418 (CISD1-Binding). For 8-Cell, n = 11, the values are 3.109 ± 0.1873 (control) and 2.703 ± 0.1292 (CISD1-Binding). For Morula, n = 12, the values are 3.457 ± 0.2498 (control) and 1.466 ± 0.2157 (CISD1-Binding). For Blastocyst group, n = 12, the values are 2.659 ± 0.1224 (control) and 2.143 ± 0.1093 (CISD1-Binding). b. Morula embryos expressed with or without CISD1-Bindingprotein (red) were stained with Sybgreen I (green) for observing mtDNA. The insets indicate that the embryo with CISD1-Binding protein expression has less mtDNA than in the control embryo. Bar = 40 μm. Three biologically repeated experiments with similar results.
