Figure 1
From: NARFL deficiency caused mitochondrial dysfunction in lung cancer cells by HIF-1α–DNMT1 axis
Knockdown NARFL caused mitochondrial dysfunction in two cell lines. (A) Knockdown assay was performed using shRNA, and the results showed that it was effective for NARFL knockdown in A549 and H1299 cells. (B–C) mtND1-6 and mtND4L mRNA levels were detected using RT-qRCR method. ND2, ND3, ND5 and ND6 was significantly downregulated in A549 cells (B) and H1299 cells (C), but increased in ND1. (D–E), Mitochondrial Complex activities were measured using activity kit, and NARFL knockdown decreased the activities of Complex I and Complex II–III, but not Complex IV and Complex V. (F) mtDNA copy number was tested and it was significantly downregulated in two cell lines treated with NARFL knockdown. (G) ATP levels were significantly reduced in two cells using NARFL knockdown. These results indicated that NARFL knockdown could affect the ND genes expressions and mitochondrial Complex activities, and it would downregulate the mitochondrial complex activity and energy generation. (H) TEM was used for observation of mitochondria. Mitochondrial membrane density was increased, but the quantity was decreased in two cells.
