Table 2.

In vivo tick dual luciferase assay.

Tick life stage	Plasmid	Luciferase cassette(s)	Relative RlucBb units/biological replicatea			Relative FlucBb units/biological replicateb
			1	2	3	1	2	3
Fed Larvaee	PBS	n/ac	85.2	89.6	92.8	20.0	24.8	24.4
	pJSB175	flaBp-flucBb	37.6	36.4	34.4	137.6	245.6	138
			LoQd = 52.3			LoQ = 49.7
	pCFA701	flaBp-rlucBb	185.6	108.8	208.0	22.4*	24.0*	24.0*
	pCFA801	flaBp-rlucBb; flaBp-flucBb	603.6	1,502.4	1,187.6	56.0	136.0	100.8
	pCFA802	flaBp-rlucBb; ospAp-flucBb	144.4	153.2	176.0	26.0*	25.2*	24.0*
	pCFA803	flaBp-rlucBb; ospCp-flucBb	346.8	341.2	234.8	22.4*	23.6*	19.6*
Fed Nymphf	PBS	n/a	82.4	93.2	83.2	19.6	20.4	18.8
	pJSB175	flaBp-flucBb	31.6	31.6	30.0	504.8	1,700.8	881.6
			LoQ = 40.3			LoQ = 27.6
	pCFA701	flaBp-rlucBb	41.6	29.6*	296.0	18.0*	19.6*	18.0*
	pCFA801	flaBp-rlucBb; flaBp-flucBb	2,416.8	2,560.0	780.4	956	552.8	388
	pCFA802	flaBp-rlucBb; ospAp-flucBb	1,001.2	2,257.6	338.4	107.6	262	36
	pCFA803	flaBp-rlucBb; ospCp-flucBb	1,500.8	1,239.6	34.8*	21.2*	32.8	16.4*

^aRelative Rluc_Bb units from three independent tick extracts incubated with 3.5 mM coelenterazine.

^bRelative Fluc_Bb units from three independent tick extracts incubated with 700 μM luciferin.

^cNot applicable.

^dLimit of Quantification (LoQ) defined as the average background signal for each assay plus 10 standard deviations.

^eExtract from groups of 24 fed larvae crushed in PBS.

^fExtract from groups of 8 fed nymphs crushed in PBS.

^*Samples that fell below their respective LoQ.