Fig. 4.

Segregation of K^b–peptide clusters is impaired when K^b is truncated of cytoplasmic tail. (A) L929 cells stably transfected with wild-type (L929/K^b) or cytoplasmic tail-deleted K^b (L929/ΔK^b) were coinfected with VV-Ub-SIIN and VV-Ub-SIYR (MOI = 1 for each). Four hours later, intracellular K^b–SIYR and K^b–SIIN complexes were costained with 2Cm67 and 25D1.16 as described in Fig. 1C. The colocalization between K^b–SIYR and K^b–SIIN complexes was analyzed with NIH Image J. The P value is 0.0006 for colocalization correlation coefficient analyzed with ∼10 images from each cell line by Mann-Whitney nonparametric two-tail analysis. (B) Cells infected as in A were stained with Alexa488-2Cm67 and Alexa647-25D1.16 Fab and imaged using dual-color live TIRF. Forty images for each condition were used for statistical analysis. The P value is less than 0.0001 using a Mann-Whitney nonparametric two-tail analysis.