Fig.1 : ― 272 ―(A) Loss of TFAM or GBA protein results in cytosolic leakage of mitochondrial DNA. (A) Quantitative PCR (qPCR) analysis of TFAM and GBA mRNA levels 3 days post-siRNA transfection. (B) Immunostaining for dsDNA, histone H2B, and Hsp60 in HeLa cells transfected with control siRNA (day 3). The “Subtract-1” image depicts the subtraction of the Hsp60 image from the dsDNA image. The “Subtract-2” image shows the subtraction of the Hsp60 image and histone-H2B image from the dsDNA image. The image on the far right is an enlargement of the red square in the “Subtract-2” image. (C) Knockdown of TFAM expression using siRNAs in HeLa cells. Immunostaining for dsDNA, histone H2B, and Hsp60 in HeLa cells transfected with TFAM siRNA (day 3). The “Subtract-1” image illustrates the subtraction of the Hsp60 image from the dsDNA image. The “Subtract-2” image displays the subtraction of the Hsp60 image and histone-H2B image from the dsDNA image. The image on the far right is an enlargement of the red square in the “Subtract-2” image. (D) Knockdown of GBA expression using siRNAs in HeLa cells. Immunostaining for dsDNA, histone H2B, and Hsp60 in HeLa cells transfected with GBA siRNA (day 3). The “Subtract-1” image indicates the subtraction of the Hsp60 image from the dsDNA image. The “Subtract-2” image exhibits the subtraction of the Hsp60 image and histone-H2B image from the dsDNA image. The image on the far right is an enlargement of the red square in the “Subtract-2” image. (E) The bar graph presents the ratio of ectopic mitochondrial DNA dots per cell number among HeLa cells transfected with control, TFAM, and GBA siRNAs.
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