Mitochondrial transcription factor A (TFAM) was knocked down in HTR-8/SVneo cells

As shown in the supplementary Figure 1 A, HTR-8/SVneo cells were transfected with negative control siRNA and three different sequences of siTFAM, respectively. Compared with the blank control group, TFAM mRNA levels in the negative control group showed no statistical difference (P = 0.467), but were significantly decreased in siTFAM #1, #2 and #3 (P < 0.001). Furthermore, TFAM protein expression decreased after transfection of siTFAM #1 (B). Moreover, ND-1/β2M and COX-3/β2M in the siTFAM group were significantly lower than those in the negative control group (P = 0.001; P < 0.001), indicating that the mtDNA copy number of trophoblast cells was significantly decreased (C, D). *P< 0.05; **P< 0.01; ***P< 0.001.