Fig. 3
PC1/3 and PC2 RNAi affect larval growth and survival. All larvae were injected at 12 days after egg lay (4th larval instar) at a concentration of 1 µg/µl unless stated otherwise. Blue circles indicate successful metamorphosis to pupae, red crosses mark deaths. A Growth curves of individual larvae injected with dsRed-dsRNA (n = 5). Larvae undergo constant growth until a weight of app. 2 mg is reached. Larvae lose weight during the pre-pupal stage and pupate between day 21 and 23 of development. One larva with transiently reduced growth pupated later but at a similar weight. These results were similar to untreated wild-type larvae (see Additional file 1: Figure S4). B PC1/3-RNAi larvae (n = 8) show strongly reduced growth and 50% die prematurely. The other half reach the pupal stage with strong delay (all surviving spent more than 35 days at the larval stage). C PC2-RNAi larvae (n = 8) initially show a better growth than PC1/3-larvae but then all suffer severe weight loss and die as larvae. Lethality is 100%. D Double PC1/3- and PC2-RNAi (1 µg/µl each) (n = 8) leads to a combined phenotype with no weight gain and early death. Total injected dsRNA concentration was 2 µg/µl, see Additional file 1: Figure S4. E Average growth curves of each RNAi treatment based on data shown in A-D and including a dsRed control injected at 2 µg/µl, matching the total dsRNA concentration of D (individual curves not shown). Error bars indicate standard deviations. Green circles indicate the time interval between two data points in which > 50% of larvae completed a moult. Dashed lines mark the interval by which the remaining (surviving) larvae completed the moult (line going to the right means additional moults occurred after the main data point, line going to left indicates additional moults have occurred before). Green circles marked with dsR1 and dsR2 on day 13 indicate that > 50% of dsRed 1 µg/µl- and 2 µg/µl-injected larvae underwent a moult between injection and first data acquisition. Black asterisks indicate that a small proportion (< 50%) of the PC2-RNAi larvae (1/8) and of the PC1/3-PC2-RNAi larvae (3/8) underwent a moult in the respective time intervals. PC1/3-RNAi larvae undergo supernumerary moults while showing severely reduced weight gain. Graph only includes development to day 42, after which no more moults occurred. Individual data of PC1/3 larval growth and moult cycle is given in Additional file 1: Table S4