Table 1.

Salivary proteins of female R. microplus ticks recognized by sera from naïve and/or two-times infested genetically tick-susceptible and/or tick-resistant bovines

Spot No. in figure	CDS in sialotranscriptomea	Annotation and putative function of protein	MW Kdab	Source of pooled reactive serac
34	20757	14-3-3 CG17870-PA, isoform A isoform 2	27	NR IR
33	20757	14-3-3 CG17870-PA, isoform A isoform 2	27	NR IR
20	18420	actin	42	IR
21	18420	actin	42	IR
19	18420	actin	42	IR IS
22	18420	actin	42	IR NS
1	109972	anticoagulant protein rhipilin-1	17	NR IR NS IS
37	31546	apolipophorin	89	NR IR
18	31546	apolipophorin	89	NR IR
22	7197	beta tubulin	53	IR NS
22	22306	beta tubulin	50	IR NS
22	22310	beta tubulin	35	IR NS
29	14491	cathepsin D2	42	NS IS NR IR
20	39114	cathepsin L-like cysteine proteinase B	38	IR
21	39114	cathepsin L-like cysteine proteinase B	38	IR
19	39114	cathepsin L-like cysteine proteinase B	38	IR IS
1	50205	CBF1-interacting corepressor	30	NR IR NS IS
40	30215	chaperonin subunit	65	IR
9	111414	CRISP3-cysteine-rich secretory protein	46	IR NS
30	25106	enolase	43	IR
24	67998	ENSANGP00000022132	25	NR IR
22	4388	F0F1-type ATP synthase, beta subunit	59	IR NS
25	111630	glutathione S-transferase	31	NR
24	111630	glutathione S-transferase	31	NR IR
38	21956	heat shock protein	71	NS
37	4142	heavy-chain filboin	44	NR IR
14	71787	hypothetical protein	22	NR IR IS
1	110513	hypothetical protein	3	NR IR NS IS
38	121267	hypothetical protein BRAFLDRAFT_287019	45	NS
20	122308	longipain cystein protease	33	IR
21	122308	longipain cystein protease	33	IR
19	122308	longipain cystein protease	33	IR IS
13	113489	lospin 8 type serpin	43	IR IS
12	113489	lospin 8 type serpin	43	NR IR IS
11	113489	lospin 8 type serpin	43	NR IR NS IS
6	113489	lospin 8 type serpin	43	NR IR NS IS
7	113489	lospin 8 type serpin	43	NR IR NS IS
10	113489	lospin 8 type serpin	43	IR
8	113489	lospin 8 type serpin	43	IR IS
5	113489	lospin 8 type serpin	43	NR IR IS
11	171727	lysosomal acid phosphatase	26	NR IR NS IS
40	9166	protein disulfite isomerase-2	39	IR
40	10534	protein disulfite isomerase-2	39	IR
12	39744	putative chitinase	45	NR IR IS
15	128399	putative salivary secreted protein	36	IR IS
24	83247	putative salivary secreted protein	25	NR IR
14	77570	putative salivary secreted protein	25	NR IR IS
14	85307	putative salivary secreted protein	27	NR IR IS
35	108605	putative secreted protein	24	IR
35	171393	putative secreted protein	18	IR
32	171393	putative secreted protein	18	NR IR
33	171393	putative secreted protein	18	NR IR
34	171393	putative secreted protein	18	NR IR
30	174664	putative secreted protein (histamine-binding?)	21	IR
31	174664	putative secreted protein (histamine-binding?)	21	NR IR
32	174664	putative secreted protein (histamine-binding?)	21	NR IR
33	174664	putative secreted protein (histamine-binding?)	21	NR IR
34	174664	putative secreted protein (histamine-binding?)	21	NR IR
31	178127	putative secreted protein (histamine-binding?)	21	NR IR
33	178127	putative secreted protein (histamine-binding?)	21	NR IR
34	178127	putative secreted protein (histamine-binding?)	21	NR IR
35	178127	putative secreted protein (histamine-binding?)	21	IR
14	54776	putative thyropin precursor	23	NR IR IS
17	113785	salivary lipid interacting protein	20	NR IR
36	113785	salivary lipid interacting protein	20	NR IR
16	113785	salivary lipid interacting protein	20	NR IR
8	38904	secreted protein	38	IR IS
31	164102	secreted protein	23	NR IR
31	164103	secreted protein	23	NR IR
33	164103	secreted protein	23	NR IR
7	38904	secreted protein	38	NR IR NS IS
16	128399	secreted protein	36	NR IR
17	128399	secreted protein	36	NR IR
26	8103	secreted salivary gland peptide	25	IS IR
25	8103	secreted salivary gland peptide	25	NR
24	8103	secreted salivary gland peptide	25	NR IR
27	8103	secreted salivary gland peptide	25	NR IR NS
24	26121	selenium-dependent salivary glutathione peroxidase	18	NR IR
20	106322	serine proteinase inhibitor serpin-3	43	IR
21	106322	serine proteinase inhibitor serpin-3	43	IR
19	106322	serine proteinase inhibitor serpin-3	43	IR IS
30	106322	serine proteinase inhibitor serpin-3	43	IR
20	106321	serine proteinase inhibitor serpin-3	43	IR
19	106321	serine proteinase inhibitor serpin-3	43	IR IS
12	6955	serpin-2 precursor	36	NR IR IS
28	13343	translation elongation factor EF-1 alpha-Tu	51	NS IS
29	13343	translation elongation factor EF-1 alpha-Tu	51	NS IS NR IR
31	32190	uncharacterized protein	17	NR IR
37	43507	vitellogenin-1	158	NR IR
18	43508	vitellogenin-1	97	NR IR
18	43509	vitellogenin-1	182	NR IR

Note: The table is formatted according to annotation of putative function of protein, but not by spot number recovered from gel. In some instances there are more than one protein identified in the spot collected

^aCDS: coding sequences generated by the sialotranscriptome of R. microplus and deposited at GenBank - NCBI (BioProject ID PRJNA329522)

^bMolecular weight furnished by Scaffold software

^c Abbreviations for sources of sera and samples: NR, pooled sera collected from four tick-naïve, genetically resistant Nelore; IR, pooled sera from four genetically resistant, twice-infested Nelore bovines at the end of the infestation; NS, pooled sera collected from four tick-naïve, genetically susceptible Holstein bovines; IS, pooled sera from four genetically susceptible, twice-infested Holstein bovines at the end of the infestation

Salivary proteins were obtained from saliva of female R. microplus ticks feeding on genetically susceptible hosts. Sera employed were obtained from naïve and/or two-times infested genetically tick-susceptible and/or tick -resistant bovines. Criteria employed to identify each sequence: minimum of one peptide presenting with 90% probability of being the protein and 90% probability of being the peptide