Fig. 3

Fig. 3

Quantification of Protein Association in the Presomitic Mesoderm

(A) A schematic of cell transplantation and FCCS data collection. For FCCS, the confocal volume (drawn approximately to scale in blue in A and B) was positioned at a membrane interface between RFP and GFP expressing cells.

(B) Confocal image of a mosaic embryo comprised of cells expressing mem-RFP and mem-GFP. The scale bar represents 10 µm.

(C) Autocorrelation and cross-correlation measurements of Itgα5-RFP and Itgα5-GFP in a mosaic interface show low cross-correlation.

(D) Plots of cross-correlation from individual measurements and mean values.

(E) Table of cross-correlation (F_cross) magnitude ± SD. The significance was examined using both t test (two tailed, unequal variance) and Mann-Whitney U. There were no statistically significant differences in mem-GFP/mem-RFP compared to Itgα5-GFP/mem-RFP and Itgα5-GFP/Itgα5-RFP compared to Itgα5-GFP/Itgα5^FYLDD-RFP. Notably, either mem-GFP/mem-RFP or Itgα5-GFP/mem-RFP compared to either Itgα5-GFP/Itgα5-RFP or Itgα5-GFP/Itgα5^FYLDD-RFP differed (p < 0.002). The Itgα5-RFP/Cdh2-GFP trans F_cross differs from the negative control (p < 0.05), but not from the Itgα5-RFP/Cdh2-GFP cis F_cross. The Itgα5-GFP/Itgα5-RFP F_cross in the absence of Cdh2 is significantly lower (p < 0.05) than the Itgα5-GFP/Itgα5-RFP F_cross in the presence of Cdh2. See Figure S1 for apparent K_D calculations.