Figure 1. Screening strategy.

(a) Cartoon summarizing the two main functions of the dynein co-factors NudE and Lis1: regulation of motor activity and recruitment of dynein to cargo. (b) Proof of concept experiments. Graph shows the number of larval progeny per adult, termed reproductive fitness (RF), in the nud-2(+) control and the null allele nud-2(ok949), after targeting genes with dsRNA using the bacterial feeding method in a 96-well liquid format. Empty dsRNA vector and hil-5(RNAi) are negative controls, plk-1(RNAi) is a control for the efficiency of RNAi, and lis-1(RNAi) is the positive control that results in enhanced lethality in nud-2(ok949) relative to nud-2(+). Error bars represent the SEM with a 95 % confidence interval, and n denotes the number of adults whose progeny were counted. (c) The RF ratio, defined as the RF of nud-2(ok949) divided by the RF of nud-2(+) for the conditions shown in (b). Error bars represent the SEM with a 95 % confidence interval. (d) Flowchart of the screening protocol. The primary screen was performed in two technical replicates per dsRNA vector and strain. The secondary screen was performed in three biological replicates with two technical replicates each. An RF ratio of 0.5 was used as a cutoff for genetic interactors of nud-2. (e) The number of progeny per well plotted against the number of parents per well for the primary genome-wide screen. For 3 - 13 parents treated with empty dsRNA vector the correlation is linear and thus suitable for evaluation of reproductive fitness. Error bars represent the SEM. (f) Example image of a screening well captured with a 2x objective. Blow up on the right shows GFP-labelled pharynxes, which can be automatically assigned to parents (green) or progeny (blue) based on their size. Scale bar, 1 mm; blow-up, 200 μm.