Table 2. Amaxa nucleofection assays.

Strain		Amaxa Solution	Program number	Vector	Positive assay/total	Transfectant analysis
						PCR	Fluo	Activity
PCF
	IL3000	T-cells	X-001, X-006, U-033	pLEW13	0/3
		Parasitic 1	U-033	pLEW13	0/1
		Parasitic 2	X-001	pLEW13	1/1	X
			X-001	pLEW20c-GFP	1/1		X
	IL3000:13	Parasitic 2	X-001	pLEW29c	1/2	X
	IL3000:13–29	Parasitic 2	X-001	pLEW20c-GFP	1/1		X
			X-001	pLEW20c-Rluc	1/1			X
			X-001	p2T7Ti/αTUB	1/2	X
	Treu183:13–29	Parasitic 2	X-001	pLEW20c-GFP	1/1		X
				pLEW20c-Rluc	1/1			X
				p2T7Ti/αTUB	1/1	X
BSF
	IL3000	T-cells	X-001, X-006	pLEW13	0/2
		Parasitic 1	U-033	pLEW13	1/1	X
		Parasitic 2	U-033	pLEW13	1/1	X
			U-033	pLEW20c-Rluc	0/1
	IL3000:13	Parasitic 2	X-001, U-033	pLEW29c	0/7
	STIB910	T-cells	X-001, U-033	pLEW13	0/3
			X-006	pLEW13	1/1	X
		Parasitic 1	X-001, X-006	pLEW13	0/2
		Parasitic 2	X-001	pLEW13	1/1	X
			X-001	pLEW20c-Rluc	1/1			X
	STIB910:13	Parasitic 2	X-001, U-033	pLEW29c	0/8
			X-001	p2T7Ti/αTUB	0/1
	IL3000:13–29	Parasitic 2	X-001	pLEW20c-Rluc	1/1			X

Once selected, transfectants were analyzed by PCR on genomic DNA, observation of EGFP fluorescence, or Renilla luciferase activity measurement.