Figure 8.

Effect of CMT2B-associated Rab7 mutations on EGFR degradation. HeLa cells were transfected for 20 h with HA-tagged Rab7 wt, Rab7 T22N, Rab7 Q67L, Rab7 L129F, Rab7 N161T, Rab7 V162M, or nontransfected (NT) as indicated. Cells were then treated with cycloheximide for 1 h and subsequently with EGF for 15 or 180 min. A, Lysates were subjected to Western blot analysis using an anti-EGFR antibody. Western blot with anti-tubulin antibody on the same membrane was used to verify equal loadings. B, The intensities of EGFR staining were quantified and plotted as a percentage of the respective intensities after 15 min of EGF stimulation. Values at 15 min were set to 100%. Values are means of four independent experiments ± SEM (error bars). Differences between the values of cells expressing Rab7 T22N and the other samples are statistically significant (p value <0.01); differences between the values of cells expressing Rab7 wt and cells expressing CMT2B associated Rab7 mutants are not statistically significant.