Fig. 2

Homology analysis and mRNA expression of HlDFS1 and HlDFS2. Sequence alignment (a) and phylogenetic tree (b) constructed with sequences of defensins from H. longicornis and other species. HlDFS1 and HlDFS2 are indicated with red asterisks. Sequence GenBank accession numbers: H. longicornis 1 (ABO28925.1); H. longicornis 2 (BAD93183.1); R. microplus (AAO48943); O. moubata (BAB41028); O. papillipes (ACJ04425.1); O. rostratus (ACJ04428.1); C. puertoricensis (ACJ04429.1); A. monolakensis (ABI52766.1); D. silvarum (AJG42673.1); I. ovatus (BAH09305.1); I. persulcatus (BAH09304.1); A. americanum (ABI74752.1); R. philippinarum (ADO32580.1); C. gigas (ACQ76287.1): D. melanogaster (AAF58855.1). c RT-PCR (upper panel) and quantitative RT-PCR (lower panel) analysis of HlDFS1 and HlDFS2 mRNA expression in unfed and fed adult H. longicornis. RNA were isolated from unfed and fed (for 72 h) whole ticks, and expression level of HlDFS1 and HlDFS2 mRNA were amplified by semi-quantitative and SYBR Green quantitative PCR using gene specific PCR (Additional file 1: Table S1). Tick actin gene was amplified as the loading control. Results are expressed as the mean ± SEM. *P < 0.05 and **P < 0.01 (t-test). The representative results from at least three independent experiments are shown