FIG 4.

Knockdown of autophagy in the female germ line decreases wMel density. Stage-specific confocal analysis reveals a decrease in relative Wolbachia density upon knockdown of autophagy genes during multiple stages of development. NGT;nos was used to knockdown Atg1, and MTD was used to knockdown Atg8a. DNA is colored cyan, D cadherin (labeling the follicle cells) is yellow, and a fluorescently conjugated DNA probe detecting Wolbachia is in magenta. Grayscale images display the Wolbachia-only channel. (A) Representative confocal z-stack images of control and Atg1 knockdown in the germ line of wMel-infected flies. (B) Quantification of wMel-infected stage-specific egg chambers upon Atg1 knockdown. Statistically significant P values are reported only. (C) Representative confocal z-stack images of wMel-infected stage-8 control egg chambers show high levels of germ line Wolbachia. (D) Representative confocal z-stack image of stage-8 egg chambers with Atg8 RNAi expression displays reduced wMel density in the germ line. (E) Quantification of wMel-infected stage-8 egg chambers for the control and Atg8 knockdown. Scale bars, 10 µM for stages 2 and 3, 20 µM for stages 4 to 6, and 40 µM for stages 7 and 8. Student’s t tests were conducted to determine significance.