Figure 8. Lpp-lipids are required for Spiroplasma proliferation.

(A) Quantification of DAG levels in hemolymph of 12-day-old flies 8 days after knockdown of Lpp by RNAi. Mean ± SEM of three independent experiments is shown, *p=0.015. (B) Quantification of sterol concentration in hemolymph of 12-day-old flies 8 days after knockdown of Lpp by RNAi. *p=0.0125. Mean ± SEM of three independent experiments is shown. (C) Spiroplasma titers quantified by qPCR in flies which have Lpp expression knocked down by RNAi relative to control flies. Spiroplasma titers were quantified at 7, 14, 21, and 28 days after activation of RNAi. Mean ± SEM of at least three samples is shown (five flies pooled per sample). ***p=0.0005 (14 days), *p<0.01 (21 days), and *p<0.01 (28 days). Shown is one representative experiment out of three independent experiments. (D) The survival of Spiroplasma-infected (Sp (+)) and Spiroplasma-uninfected (Sp (−)) flies with or without RNAi-mediated Lpp knocked down. ***p<0.0001, N = 18 flies per condition. Shown is one representative experiment out of three independent experiments.

DOI: http://dx.doi.org/10.7554/eLife.02964.017

Figure 8—figure supplement 1. Hemolymph protein is not decreased by Lpp depletion.

Protein concentration in the hemolymph of flies that are 12 days old, 8 days after activation of Lpp-RNAi. NS (p=0.908). Mean ± SEM of three independent experiments is shown.

DOI: http://dx.doi.org/10.7554/eLife.02964.018

Figure 8—figure supplement 2. Spiroplasma titers are decreased by Lpp depletion.

(A) Spiroplasma titers quantified by qPCR in flies which have Lpp-RNAi construct which has not been activated by heat shock relative to control flies without the Lpp-RNAi construct, NS (p=0.587), and an additional control where Lpp-RNAi has been activated, **p=0.0037. Spiroplasma titers were quantified at 14 days of fly age. Mean ± SEM of at least three samples is shown (five flies pooled per sample). Shown is one representative experiment out of the three independent experiments. (B) Spiroplasma titers quantified by qPCR in flies in which Lpp has been knocked down by the expression of the UAS-LppRNAi(46) using the C564>gal4^TS driver. Flies were raised at the restrictive temperature of 18°C where C564>gal4 activity is repressed by gal80ts and then shifted to 29°C at 4 days post-eclosion (Lpp-RNAi is then activated) where they were maintained for 25 days. *p=0.0215. Mean ± SEM of at least three samples is shown (five flies pooled per sample). Shown is one representative experiment out of the three independent experiments.

DOI: http://dx.doi.org/10.7554/eLife.02964.019