TABLE 1:

FCS measurements of secreted fractions of HhGFP.

Sample	D1 (μm2/s)	Fraction (%)	D2 (μm2/s)	Fraction (%)
HhGFP P100	40.1 (± 3.78)	79.5 (± 4.9)	5.40 (± 1.40)	20.4 (± 4.9)
HhGFP P250	40.8 (± 2.93)	94.9 (± 3.5)	5.66 (± 1.74)	5.1 (± 3.5)
HhGFP S250	85.5 (± 15.2)	97.3 (± 3.1)	5.52 (± 3.81)	2.7 (± 3.1)
GFP solution	83.7 (± 3.77)	5.1 (± 2.8)

Diffusion coefficients and fractions of individual components obtained after fitting the FCS data to a two- component, 3D diffusion fit. Although the diffusion coefficients of two forms are similar in both fractions, there is a significant reduction in the fraction of the slow-diffusing component in P250 for both HhGFPs. The diffusion coefficient of the S250 form matches that of GFP in solution (last row), indicating the presence of either a monomeric or a dimeric species in the S250 form.