Additional file 1: Figure S1. of Thioester-containing proteins regulate the Toll pathway and play a role in Drosophila defence against microbial pathogens and parasitoid wasps
Creators: name: Dostálová, Anna
name: Rommelaere, Samuel
name: Poidevin, Mickael
name: Lemaitre, Bruno
Publisher: name: Figshare
Year: 2017
Subjects: subject: Microbiology
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: Cell Biology
subject: Genetics
subject: Molecular Biology
subject: Neuroscience
subject: Physiology
subject: Biotechnology
subject: Evolutionary Biology
subject: 39999 Chemical Sciences not elsewhere classified
subject: FOS: Chemical sciences
subject: FOS: Chemical sciences
subject: Ecology
subject: Immunology
subject: FOS: Clinical medicine
subject: FOS: Clinical medicine
subject: Marine Biology
subject: Cancer
subject: Infectious Diseases
subject: FOS: Health sciences
subject: FOS: Health sciences

description: A. Molecular confirmation of the TEPq Δ mutant. Expression of TEP1, 2, 3 and 4 was monitored by qRT-PCR in the w 1118 and TEPq Δ flies after bacterial challenge. Female flies were pricked in the thorax with a needle dipped in a concentrated mixed culture of E. faecalis (OD 0.5) and E. carotovora (OD 200). Expression of TEP genes was measured 6 h post-infection using primers TEP1F, TEP1R, TEP2F, TEP2R, TEP3F, TEP3R, TEP4F and TEP4R (for sequences see Additional file 3: Table S1). The expression of all four genes was strongly reduced in the TEPq Δ line as compared to w 1118 , confirming the TEPq Δ genotype. Furthermore, we confirmed the presence of the expected transposon insertion (see Fig. 1b) in the TEPq Δ mutant in the respective loci by PCR. Insertion of the MiMiC element in the Tep1 locus was confirmed using primers dTEP1F and dTEP1R. An approximately 700-bp fragment was amplified in the TEPq Δ mutant but not in the control wild-type line. Insertion of the EP element in the Tep4 locus was confirmed using primers dTEP4F and dTEP4R. An approximately 330-bp fragment was amplified in the TEPq Δ mutant but not in the control wild-type line. Presence of a deletion due to the flippase excision of the genomic region between the flippase recognition target (FRT) sites of the two XP elements in the Tep2 and Tep3 loci was confirmed using primers dTEP2&amp;3 F and dTEP2&amp;3R. No amplification was observed in the TEPq Δ mutant; an approximately 630-bp fragment was amplified in the control wild-type line. Sequences of all primers used are indicated in Additional file 3: Table S1. B. Viability of the TEPq Δ line. TEPq Δ /CyO males were crossed to TEPq Δ homozygous females, and the ratio of homozygous versus heterozygous flies was counted. A significantly higher number of TEPq Δ /CyO offspring was observed (P  0.05). Shown is the level of Socs36E expression normalised to RpL32 in unchallenged (UC) flies and in flies collected 2 h after being pricked in the thorax with a clean needle. D. Flies were pricked in the thorax with a clean needle, and the level of melanisation at the wound site, estimated by the size of the melanin spot, was examined 3 h later. TEPq Δ flies showed a normal rate of surface melanisation of the wound site. E. Melanisation in flies after infection with P. luminescens. No statistically significant difference was observed between the TEPq Δ , TEP4 Δ and wild-type flies (chi-square test, P = 0.0681). (PDF 526 kb)