<i>fard-1</i> RNAi Treatments Results in Compromised Ether-Linked Lipid Abundance.

<p>(A) Developmental RNAi-treatment was initiated at the L1 stage and continued until animals were harvested for analysis at day 3 of adulthood, resulting in 4 days of RNAi feeding at 25°C (blue). The second RNAi-treatment described is an adult-only <i>fard-1</i> RNAi strategy (yellow) where RNAi is started after the completion of development at day 1 of adulthood (44 hours post-hatch). Animals were harvested at day 3 for both treatments. (B) qRT-PCR of <i>fard-1</i> expression is reduced to 0.34 ± 0.18 and 0.40 ± 0.17 of wild-type levels with developmental RNAi and adult-only RNAi treatment, respectively. The <i>cdc-42</i> housekeeping gene was used for normalization. (C) When <i>fard-1</i> RNAi is initiated at the L1 stage and continued throughout development until day 3 of adulthood (blue), there is a significant reduction in the relative abundance of both O-PE and P-PE compared to control L4440 RNAi (black). (D) The overall composition of PE was compared in control RNAi- (grey) and <i>fard-1</i> RNAi-treated animals (light blue) after binning the lipids by the overall degree of unsaturation found in the associated fatty acids. Similarly, PC species were measured in RNAi controls (black) and <i>fard-1</i> RNAi-treated animals (dark blue). (E) <i>fard-1</i> RNAi initiated in adult animals at day 1 and maintained until day 3 of adulthood (yellow) resulted in a significant decrease in O-PE as quantified by HPLC-MS/MS. There is a small but not significant decrease in the P-PE population (p-Value of 0.11). (F) The impact of adult-only <i>fard-1</i> RNAi treatment on the PE and PC populations showed no significant changes in the degree of unsaturation in PE (yellow) or PC (orange) compared to L4440 RNAi shown in grey for PE and black for PC. Data was generated from at least 3 independent biological replicates with SEM shown. See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0167229#pone.0167229.s006" target="_blank">S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0167229#pone.0167229.s007" target="_blank">S2</a> Datasets for complete list of phospholipid species. *p<0.05 was determined by unpaired t-tests using Holm-Sidak corrections for multiple comparisons.