Skip to main content
. 2014 Dec 15;7(1):78–101. doi: 10.15252/emmm.201404392

Figure 7. In vivo repair of mitochondrial DNA damage by HMGB1.

Figure 7

Mitochondrial DNA damage repair was evaluated in vivo in mice subjected to X-ray irradiation (20 Gy) under anaesthesia. Cerebellar tissues were collected 10, 180 and 300 min after irradiation.

  1. The mitochondrial DNA amplification assay in the cerebellar tissue obtained from wild-type (WT), Atxn1-KI (KI) and Atxn1-KI;HMGB1-Tg double-transgenic (KI+H) mice. Amplification of the long and short fragments from the mitochondrial genome was performed at each time point after irradiation.
  2. Quantitative analysis of mitochondrial DNA damage at each time point using the ratio between short and long PCR fragments (long/short). Reduction of the ratio indicates enhanced DNA damage. The data are presented as mean ± SD. #P < 0.05 in one-way ANOVA followed by post hoc Tukey's test.
  3. Reversal of mitochondrial DNA damage from 10 min to 180 min or 300 min was evaluated by subtraction of the values. The data are presented as mean ± SD. #P < 0.05 in one-way ANOVA followed by post hoc Tukey's test.
  4. The nuclear DNA amplification assay in the cerebellar tissue obtained from wild-type (WT), Atxn1-KI (KI) and Atxn1-KI;HMGB1-Tg double-transgenic (KI+H) mice. Amplification of the long and short fragments from the nuclear genome was performed at each time point after the irradiation; #P < 0.05 in one-way ANOVA followed by post hoc Tukey's test.
  5. Quantitative analysis of nuclear DNA damage at each time point using the ratio between short and long PCR fragments (long/short). A reduction in the ratio indicates enhanced DNA damage. The data are presented as mean ± SD. #P < 0.05 in one-way ANOVA followed by post hoc Tukey's test.
  6. Reversal of nuclear DNA damage from 10 to 180 min or 300 min was evaluated by subtraction of the values. The data are presented as mean ± SD. #P < 0.05 in one-way ANOVA followed by post hoc Tukey's test.
  7. Southern blot analysis of mitochondrial genomic DNA prepared from wild-type (WT), Atxn1-KI or Atxn1-KI;HMGB1-Tg double-transgenic mice. Mitochondrial DNA copy numbers were largely similar among the genotypes, although quality of the mitochondrial DNA was remarkably different.
  8. Signal intensity of a 16-kb band corresponding to the intact mitochondrial genome and those of a DNA smear (< 16 kb) corresponding to a degraded mitochondrial genome were quantified using ImageQuant LAS500 (GE Healthcare Life Sciences, Little Chalfont, UK) with ImageQuant TL software.

Source data are available online for this figure.