Fig. 2
Knockdown of vit-6 increases the ROS stress inside the nematodes. Staining of nematodes using CM-H2XRos demonstrates that ROS stress is increased by zmp-2 RNAi and even more by additional vit-6 RNAi (a). Protein carbonyls were detected by the use of an OxyBlot™ Protein Oxidation Detection Kit (b). Therefore, DNP-derivatized proteins were quantified densitometrically using β-actin as reference. Detection of Amplex Ultra Red fluorescence in the medium subsequent to the incubation of the N2 vector control or nematodes treated with either zmp-2 RNAi or zmp-2 plus vit-6 RNAi revealed an increased ROS secretion in response to both RNAi treatments versus the control (c). Using the ROS scavenger ascorbic acid at a concentration of 100 µM was able to prevent the increase in extracellular ROS due to zmp-2 RNAi (c) and also the reduction of thermotolerance caused by zmp-2 RNAi in wild-type nematodes (d). *p < 0.05; **p < 0.01; *p < 0.001 versus the N2 control