Knockdown of <i>wfs1</i> in both neurons and glial cells significantly shortens the lifespan of flies and exaggerates behavioral deficits and neurodegeneration.

<p>(<b>A</b>) Knockdown of <i>wfs1</i> in both neurons and glial cells, (<b>D</b>) but not in glial cells alone, caused age-dependent locomotor deficits as revealed by climbing assay. Average percentages of flies that climbed to the top (white), climbed to the middle (light gray), or stayed at the bottom (dark gray) of the vials. Percentages of flies that stayed at the bottom were subjected to statistical analyses. n = 3–5 independent experiments, *<i>p</i> < 0.05 by Student’s <i>t</i>-test. (<b>B</b>) Knockdown of <i>wfs1</i> in both neurons and glial cells, (<b>E</b>) but not in glial cells alone, caused neurodegeneration in the central neuropil or optic lobes of fly brain. Representative images show the central neuropil and optic lobe in paraffin-embedded brain section with hematoxylin and eosin (HE) staining from 30-day-old flies. Scale bars: 200 μm. Percentages of vacuole areas (indicated by arrowheads in the images) in central neuropil or optic lobes are shown. n = 14–19 hemispheres, ***<i>p</i> < 0.001 by Student’s <i>t</i>-test. (<b>C</b>) Knockdown of <i>wfs1</i> in both neurons and glial cells significantly shortened lifespan of flies (n = 199, <i>wfs1</i> RNAi group or 331, control group). (<b>F</b>) Knockdown of <i>wfs1</i> in glial cells did not affect lifespan up to 30 days (n = 114, <i>wfs1</i> RNAi group or 139, control group). The lifespans of flies were determined by Kaplan-Meier survival analysis with log-rank test and statistical significance was indicated in the figure. Genotypes and ages of flies are described in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1007196#pgen.1007196.s001" target="_blank">S1 Table</a>.