TABLE 1.

FLIM-FRET measurements showing a pseudo-TuSC interaction between GCP4 and GCP5

NA, no acceptor; bold indicates significant FRET efficiency.

Donor	Acceptor	τa	S.E.	Δτb	nc	FRETd	p valuee
		ns		ps		%
GCP3-GFP	NA	2.414	0.006		121
	mCherry	2.325	0.019	89	29	3.7	2.5 × 10−8
	GCP2-mC	1.981	0.026	433	60	17.9	2.4 × 10−51
	GCP3-mC	2.293	0.011	121	128	5.0	1.4 × 10−19
GCP4-GFP	NA	2.444	0.006		123
	GCP4-mC	2.391	0.008	53	97	2.2	2.5 × 10−7
	GCP5-mC	2.251	0.013	193	88	7.9	3.6 × 10−33
	GCP6-mC	2.347	0.010	97	90	4.0	3.6 × 10−2
GCP5-GFP	NA	2.425	0.008		136
	GCP4-mC	2.213	0.015	211	80	8.7	9.6 × 10−29
	GCP5-mC	2.369	0.018	55	50	2.3	2.0 × 10−3
	GCP6-mC	2.345	0.018	80	57	3.3	1.0 × 10−5
GCP6-GFP	NA	2.436	0.007		152
	GCP4-mC	2.422	0.014	13	60	0.6	1.6 × 10−1
	GCP5-mC	2.353	0.008	83	60	3.4	1.6 × 10−1
	GCP6-mC	2.354	0.012	82	44	3.4	1.7 × 10−7
GFP-GCP4	NA	2.402	0.009		60
	GCP5-mC	2.390	0.008	12	60	0.5	3.4 × 10−1
GFP-GCP5	NA	2.409	0.022		70
	GCP4-mC	2.326	0.022	83	43	3.5	2.3 × 10−2
GFP-GCP4	NA	2.499	0.008		60
	mC-GCP4	2.408	0.009	91	60	3.6	7.4 × 10−12
	mC-GCP5	2.262	0.009	237	60	9.5	3.3 × 10−37
GFP-GCP5	NA	2.460	0.009		60
	mC-GCP4	2.162	0.018	298	60	12.1	4.6 × 10−29
	mC-GCP5	2.388	0.010	72	60	2.9	3.5 × 10−7
N45C-GFP	NA	2.473	0.009		80
	GCP4-mC	2.377	0.009	96	60	3.9	5.5 × 10−11
	GCP5-mC	2.258	0.012	215	90	8.7	3.3 × 10−30
N54C-GFP	NA	2.456	0.007		120
	GCP4-mC	2.261	0.014	195	60	7.9	5.2 × 10−32
	GCP5-mC	2.412	0.010	44	60	1.8	3.2 × 10−4
GCP4(F522R-F626R)-GFP	NA	2.407	0.008		60
	GCP5-mC	2.169	0.019	238	60	9.9	1.5 × 10−20

^a Mean lifetime (in ns).

^b Δτ = τ_D − τ_DA (in ps).

^c Total number of measured centrosomes.

^d Percentage of FRET efficiency: E = 1 − (τ_DA/τ_D).

^e p value of the difference between the donor lifetimes in the presence and absence of acceptor (Student's t test).