TABLE 2.

SW-FCCS analysis of Cdc42-CRIB, -N-WASP, and -IRSp53 pairs The table summarizes data derived from SW-FCCS experiments shown in Figs. 4 and 5 on Cdc42 interacting with its effectors. The first column gives the concentration of individual proteins. The second column gives the diffusion times of proteins (see supplemental material). The third column gives percentage of complex formed. The fourth column gives estimated K_D. SW-FCCS data are averages ± S.D. (n = 7–28). K_D data are expressed as averages ± S.E. (n = 7–28). n = 7–28 cell measurements for each set of experiments.

Constructs	Concentration ± S.D.	Diffusion times (τD) ± S.D.	Complex ± S.D.	∼KD ± S.E. (R2 value)
	nm	ms	%	nm
Positive control
mRFP-GFP (tandem fusion)			32.3 ± 3.7	NDa
GFP	269.6 ± 130.0	0.8 ± 0.1
mRFP	106.6 ± 64.8	1.1 ± 0.2
Negative control
Cyto-GFP/mRFP			3.7 ± 3.4	Failed to fit linear regression
GFP	121.0 ± 45.4	0.6 ± 0.1
mRFP	78.5 ± 35.3	0.5 ± 0.1
GFP-CRIB(domain) and mRFP-Cdc42V12			51.6 ± 20.0	250 ± 25 (0.56)
GFP-CRIB	365.0 ± 206.0	147.3 ± 180.6
mRFP-Cdc42V12	177.0 ± 69.3	88.5 ± 51.1
GFP-CRIB (domain) and mRFP-Cdc42N17			5.0 ± 4.8	Failed to fit linear regression
GFP-CRIB	191.0 ± 139.0	1.3 ± 0.8
mRFP-Cdc42N17	512.0 ± 288.0	10.7 ± 0.7
GFP-Cdc42V12 and mRFP-NWASP			84.8 ± 16.2	27 ± 3 (0.44)
GFP-Cdc42V12	81.8 ± 60.6	179.0 ± 270.0
mRFP-NWASP	252.0 ± 237.0	103.9 ± 47.3
GFP-Cdc42N17 and mRFP-NWASP			8.6 ± 4.8	Failed to fit linear regression
GFP-Cdc42N17	71.6 ± 84.8	71.0 ± 76.2
mRFP-NWASP	162.0 ± 249.0	11.2 ± 15.9
GFP-Cdc42V12 and mRFP-IRSp53			30.9 ± 19.4	391 ± 33 (0.8)
GFP-Cdc42V12	76.3 ± 69.8	15.0 ± 9.2
mRFP-IRSp53	224.0 ± 161.0	8.5 ± 5.8
GFP-Cdc42N17 and mRFP-IRSp53			10.8 ± 8.8	2180 ± 494 (0.09)
GFP-Cdc42N17	221.0 ± 168.0	86.9 ± 154.1
mRFP-IRSp53	145.0 ± 143.0	7.6 ± 4.2

^aND, not determined