Skip to main content
. 2023 Apr 12;37(5):e22886. doi: 10.1096/fj.202202119R

FIGURE 6.

FIGURE 6

Vimentin distribution rescued by direct binding of vimentin filaments to kinesin‐1. Rescue experiment was done in RPE mEOS_vim (control and GAN KO) cells. Control (A, B) and GAN KO (C, D) cells treated with KIF5B (1‐560)‐scFV GCN4‐HA tag plasmid alone. Rescue condition (E, F), GAN KO cells received both vimentin‐GCN4‐flag tag and KIF5B (1‐560)‐scFV GCN4‐HA tag plasmid. mEOS_vim appeared normally distributed in control (A) and aggregated in GAN KO (C). Distribution of vimentin in control cells indicates that the expression of KIF5B (1‐560)‐scFV GCN4‐HA tag plasmid has no non‐specific effects. In rescue condition (E), mEOS_vim redistributed and stretched to the cell periphery besides the vimentin aggregate in the center. Filaments distribution was analyzed using Object Intensity Distribution module in CellProfiler. Cell was divided radially from the center to periphery and fluorescence intensity of mEOS_vim in each zone was calculated. (G) Intensity in the cell periphery was compared among the conditions. Rescue cells showed significantly higher vimentin filaments compared to the GAN KO condition. Statistical significance was determined using one‐way ANOVA and Tukey's multiple comparisons test (n = 15 cells). **p < .01; ****p < .0001.