Additional file 1 of The ecdysteroid receptor regulates salivary gland degeneration through apoptosis in Rhipicephalus haemaphysaloides

Additional file 1: Figure S1. Alignment of the deduced amino acid sequences of Rhipicephalus haemaphysaloides-EcR; Amblyomma americanum-EcR; Ixodes scapularis-EcR; Ornithodoros moubata-EcR; Bombyx mori-EcR; Drosophila melanogaster-EcR; Danio rerio-LXRα; Homo sapiens-LXRα; Mus musculus-LXRβ. All species had a DBD domain and an LBD domain, and the DBD domain had two conserved C4-type zinc finger motifs. Especially, the position of four cysteine residues in the ZF_C4 was highly conservative. In arthropods, the DBD and LBD domains were relatively conserved. Figure S2. Alignment of the deduced amino acid sequences of Rhipicephalus haemaphysaloides USP; Amblyomma americanum-USP; Ixodes scapularis-USP; Ornithodoros moubata-USP; Bombyx mori-USP; Drosophila melanogaster-USP; Danio rerio-USP; Homo sapiens-RXR; Mus musculus-RXR. Invertebrates and vertebrates both had a DBD domain and an LBD domain as EcR. Moreover, the DBD domain of USP was highly conserved not only in invertebrates but also in vertebrates, which was different from EcR. Figure S3. Phylogenetic tree constructed using amino acid sequences of the ecdysteroid receptor heterodimeric complex, EcR (a) and USP (b) of Rhipicephalus haemaphysaloides. Figure S4. Results of a BLAST search of RhEcR and RhUSP sequences against genomic sequences of six tick species. The query was RhEcR and RhUSP gene nucleotide sequences in a search against genomic sequences of six tick species (Ixodes persulcatus, Haemaphysalis longicornis, Dermacentor silvarum, Hyalomma asiaticum, Rhipicephalus sanguineus, Rhipicephalus microplus and Ixodes scapularisis). Figure S5. Temporal and spatial distribution of RhEcR and RhUSP in Rhipicephalus haemaphysaloides. Tissues were dissected from female ticks. Using qRT-PCR, relative RNA levels of RhEcR and RhUSP were measured at each time point. a In the period of blood-feeding, the expression of RhEcR2 was higher than RhEcR1 before E 24 h. After E 24 h, the expression of RhEcR1 was higher than RhEcR2. b Two isoforms of RhUSP were detected during all time points, but the level of RhUSP1 was high and RhUSP3 was very low. The data represent the mean ± SD of the experiments (10 ticks/time point) performed in triplicate and normalized to EF1α. c The translation of RhEcR isoforms was detected with anti-mouse RhEcR polyclonal antibody. After E 24 h, the expression of all isoforms of RhEcR gradually decreased. Table S1 Oligonucleotide sequences and names of the primers used for qRT-PCR, RNAi and cloning of Rhipicephalus haemaphysaloides genes. Table S2 Accession numbers for sequences of EcR/LXR and USP/RXR in vertebrates and invertebrates.