Fig. 5

Overexpression of ROMO1 disrupts embryo development by inducing mitochondrial fragmentation. A The d-2 embryo morphologies of ROMO1 OE and control groups. Scale bars, 100 μm. B Development and fragmentation rates at 2- and 4-cell stages. C Development rate of blastocyst embryos in ROMO1 KD, ROMO1 OE, ROMO1 KD + OE, and control groups. D Western blot image of ROMO1 levels in control, ROMO1 KD, ROMO1 OE, and ROMO1 KD + OE groups. **P < 0.01 indicates significant differences between treatment groups. E Relative mitochondrial DNA copy number in control, ROMO1 KD, ROMO1 OE, and ROMO1 KD + OE blastocysts. F ATP level in control, ROMO1 KD, ROMO1 OE, and ROMO1 KD + OE blastocysts. G Representative confocal images of control and ROMO1 OE blastocysts showing well-preserved mitochondrial clusters in control blastocysts but fragmented mitochondria clusters in ROMO1 OE blastocysts. Scale bars, 20 μm. H, I Quantification of confocal images of control (n = 5) and ROMO1 OE (n = 5) blastocysts showing mitochondrial fragmentation rate and relative mitochondrial cluster length. *P < 0.05, **P < 0.01, and ***P < 0.001 indicate significant differences between treatment groups