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Figure 4. Reabsorption of trehalose in BPH. (A) Experimental scheme of hemolymph and honeydew analysis. RNAi techniques and quantification of sugars in hemolymph and honeydew are described in the Appendix. Briefly, BPH female adults were injected with dsRNA solution in the segment between the thorax and abdomen and incubated for 24 h to suppress gene expression. Each EGFP1 or Nlst8 RNAi BPH was then injected with 50 nl of water or trehalose in the same segment and incubated for 24 h in a Parafilm sachet. Finally, sugars in BPH hemolymph or in honeydew were analyzed by using HPLC. Sugar contents are represented as percentages (mg/100 μl). (B) Effect of RNAi on the gene expression levels of Nlst8 at 48 h after injection. EGFP1-dsRNA served as a negative control. Error bars represent the standard error (n = 3). (C) Hemolymph sugar contents following injection of water or 1 M trehalose into EGFP1 (upper) or Nlst8 (lower) RNAi BPH. Sugar contents were analyzed at 30 min after injection (left), and at 24 h after injection (right). (D) Quantifications of trehalose in BPH honeydew. Statistical analyses were performed by using the Kruskal–Wallis test before Dunn’s multiple comparison tests. The asterisk indicates a significant difference (P = 0.0023). Image published in: Kikuta S et al. (2012) Copyright © 2012 Kikuta, Hagiwara-Komoda, Noda and Kikawada. Creative Commons Attribution license Larger Image Printer Friendly View

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