Figure 5
(A,B) Graphical transmembrane organization of mAChRs, using the Protter 1.0 software (https://wlab.ethz.ch/protter/start/). Red color in (A) indicates the signal peptide and green indicates the N-linked glycosylation motifs, blue numbers show transmembrane helices. (C) Phylogenetic relationship between vertebrate and arthropod mAChRs including mAChR-A and mAChR-B from I. scapularis/I. ricinus. D1 D1 dopamine receptor, InvD1L invertebrate-specific D1-like dopamine receptor, 5-HT receptor 5 hydroxytryptamine (serotonin) G-protein coupled receptor, D2 D2 dopamine receptor. GenBank Accession numbers are: Homo sapiens mAChR M1, NP_000729.2; Mus musculus mAChR M1, NP_001106167.1; H. sapiens mAChR M5, NP_001307846.1; M. musculus mAChR M5, NP_991352.2; H. sapiens mAChR M3, NP_000731.1; M. musculus mAChR M3, NP_150372.1; H. sapiens mAChR M2, NP_000730.1; M. musculus mAChR M2, NP_987076.2; M. musculus mAChR M4, XP_006498717.1; H. sapiens M4, NP_000732.2; I. scapularis/I. ricinus mAChR-A, XM_002403091.2; Apis mellifera mAChR-A, XM_395760.7; D. melanogaster mAChR-A, JQ922421.1; T. castaneum mAChR-A, JX174094.1; I. scapularis/I. ricinus mAChR-B, XM_002416115.2; D. melanogaster mAChR-B, JX028235.1; A. mellifera mAChR-B, XM_006558358.3; T. castaneum mAChR-B, XM_015985417.1; I. scapularis D1, XM_029975666.1; A. mellifera D1, NM_001011595.1; D. melanogaster D1, NM_001260163.1; I. scapularis 5-HT2, EEC03313.1; Bombyx mori 5-HT1, KM236101.1; A. mellifera 5-HT1, NP_001164579.1; I. scapularis InvD1L, XM_002399612.2; Anopheles gambiae InvD1L, KU948225.1; A. mellifera InvD1L, NM_001011567.1; I. scapularis D2, XM_029969460.1; A. mellifera D2, NM_001014983.1; D. melanogaster D2, NM_001031909.2. The phylogenetic three in C was constructed using neighbor-joining method in MEGA7 software41, with 500 bootstrap replications. (D) Tissue-specific PCR of machr-a and machr-b in different tissues from partially-fed I. ricinus females. Full-length gels including amplicon sizes can be found in Supplementary Fig. S11. rps4 ribosomal protein S4 transcript, SGs salivary glands, S synganglia, MT Malpighian tubules, In intestines (midguts), Tr tracheas, Ov ovaries, Do dorsa, Ca carcasses, C control (without template).
