Fig. 2: Exposure of w¹¹¹⁸ flies to rotenone causes rapid loss of dopaminergic neurons in aged flies accompanied by increased dSarm expression.

a Schematic representation of experimental setup for rotenone treatment in the aged (20-days old) flies. b, c Expression of dSarm in young flies (1-day old) exposed to 200 μM of rotenone for 1-day, 10-days, 20-days, and 30-days (b) and aged flies (20-days old) to 200 μM of rotenone for 3-days and 5-days post-treatment (n = 5) (c). d Multifocal confocal images of dopaminergic neurons following tyrosine hydroxylase (TH) immunostaining (green) and Elav immunostaining (red) in the brains of flies exposed to 200 μM rotenone at 20-days post-eclosion for 3-days (right-hand panel). Results were compared to age-matched rotenone untreated flies (left-hand panel). Scale bars = 200 μm. e Bar graph represents the number of dopaminergic neuron cell bodies in the control and rotenone (200 μM) treated fly brains (n = 3). f Representative western blot analysis of the dopaminergic neuronal marker tyrosine hydroxylase in 20-day old flies exposed to 200 μM rotenone and analyzed at day-3 post-exposure. β-actin served as a loading control and data was compared with age-matched untreated control flies (n = 3). *p < 0.05, **p < 0.01 and ***p < 0.001 compared to control flies