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. Author manuscript; available in PMC: 2014 Sep 1.
Published in final edited form as: Nat Neurosci. 2014 Jan 26;17(3):357–366. doi: 10.1038/nn.3639

Figure 3.

Figure 3

Oxidative stress causes DNA damage and heterochromatin loss in Drosophila. pH2Av and elav staining in neurons of control and Trxr-1481 hemizygous (a) or Sod2Δ02 heterozygous (b) mutant flies, n = 3 brains. Arrows indicate pH2Av positive nuclei. Scale bar is 10 μm. Quantification of comet tails in Trxr-1481 hemizygous (c) and Sod2Δ02 heterozygous (d) mutant flies compared to control, n = 6 brains (for c, **p = 0.01, t = 9.2 for two degrees of freedom, for d, **p = 0.01, t = 10.84 for two degrees of freedom, unpaired t-test). (e) H3K9me2 and HP1α levels in homogenates from heads of flies harboring loss-of-function mutations in Trxr-1 or Sod2. Full-length blots are shown in Supplementary Figure 5d. (f) Quantification of experiments in e, n = 3 heads (for H3K9me2, *p = 0.02, F = 7.98 for two degrees of freedom, for HP1α, *p = 0.03, F = 6.5 for two degrees of freedom, one-way ANOVA). Quantification of BL2 reporter activation in brains of Trxr-1481 (g) and Sod2Δ02 (h) flies; control is hemizygous for the BL2 reporter. n = 6 brains (for g, **p < 0.001, t = 20.19 for five degrees of freedom, for h, **p < 0.001, t = 26.56 for five degrees of freedom, unpaired t-test). (i) H3K9me2 and HP1α levels in homogenates from heads of flies with neuronal overexpression of CycA or Rheb. Full-length blots are shown in Supplementary Figure 5e. (j) Quantification of experiments in i, n = 3 heads (for H3K9me2, p = 0.92, F = 0.08 for two degrees of freedom, for HP1α, p = 0.54, F = 0.71 for two degrees of freedom, one-way ANOVA). (k) Cell cycle activation assayed by PCNA staining in brains of flies with RNAi-mediated knockdown of Su(var)205 or Su(var)3-9. n = 6 brains, **p < 0.001, F = 23.52 for two degrees of freedom, one-way ANOVA. Control is w1118 in a-h, and elav-GAL4/+ in i-k. Trxr-1481 hemizygous flies and respective controls are 2 days old, all other flies are 10 days old. Data are mean ± s.e.m., except in c, d, g and h in which data are represented as box and whisker plots comprised of minimum, lower quartile, median, upper quartile and maximum values.