Emma Warr and Suchismita Das contributed equally to this study.
The Gram-Negative Bacteria-Binding Protein gene family: Its role in the innate immune system of Anopheles gambiae and in anti-Plasmodium defence
Article first published online: 30 JAN 2008
© 2008 The Authors
Insect Molecular Biology
Volume 17, Issue 1, pages 39–51, February 2008
How to Cite
Warr, E., Das, S., Dong, Y. and Dimopoulos, G. (2008), The Gram-Negative Bacteria-Binding Protein gene family: Its role in the innate immune system of Anopheles gambiae and in anti-Plasmodium defence. Insect Molecular Biology, 17: 39–51. doi: 10.1111/j.1365-2583.2008.00778.x
Present address: Liverpool School of Tropical Medicine, Pembroke Place, Liverpool, L3 5QA. UK.
- Issue published online: 30 JAN 2008
- Article first published online: 30 JAN 2008
- Received 11 September 2007; accepted after revision 4 October 2007.
Table S1. T7 GNBP primers used for dsRNA synthesis; 1 denotes the forward primer and 2 the reverse primer. The first 18 bases of each primer sequence (TAATACGACTCACTATAG) correspond to the T7 polymerase promoter site.
Table S2. Primers used for RNAi verification (Fig. 3C) by qRT-PCR. Some new primers were designed, and the others were from the dsRNA synthesis primer set (marked as RNAI).
Table S3. Microarray log2 values of the six A. gambiae GNBPs in various tissues (midgut versus carcass and cardia versus posterior gut) and in whole mosquitoes exposed to various microbial challenges (B. subtilis, E. coli, S. typhimurium, and S. aureus). For each kind of array, three biological replicates were done.
Table S4. Efficiency of GNBP knock-down in A. gambiae female mosquitoes after dsRNA injections. RNA was extracted from both the GNBP- and GFP-silenced mosquitoes, and the relative mRNA level was determined for each gene. The mRNA level in GFP mosquitoes was taken as 1, and the corresponding % silencing was determined by qRT-PCR. The standard error (S.E.) is also shown.
Table S5. Effect of A. gambiae GNBP gene silencing on P. falciparum and P. berghei infections (oocyst numbers). The gene that has been silenced and the number of midguts dissected are shown for each kind of infection. The mean oocyst number (± the standard error), along with the oocysts range and the corresponding p-value (from the Mann-Whitney test), are also shown. The -fold difference in infection with respect to the GFP control, and the significance (S) of the -fold difference in infection level are also shown. The prevalence is the percentage of mosquitoes with at least one oocyst in the midgut.
Table S6. Regulation of eight immune genes (taken from Meister et al., 2005) after Gram-positive and Gram-negative challenges in GNBPB4-silenced Sua5B cells. De-logged ratios for each gene, before and after challenge, were determined by qRT-PCR. A ratio <0.5 indicates a down-regulation of two-fold or greater with the challenge. The transcript ID numbers (from ENSEMBL), the standard deviation (S.D.), and the standard error (S.E.) are shown for each genes for the two challenges.
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