Figure 7.

Cycloheximide treatment does not block localization of E-cadherin to intracellular vesicles. Keratinocytes cultured in normal growth medium were treated with cycloheximide to block protein synthesis (B) or left untreated (A) and switched to low-calcium medium for 4 h to induce vesicles. Cells were fixed and stained for E-cadherin expression. Images are from 0.4-μm sections taken 4.4 μm from basal surface of the cell. Scale bar, 10 μm. To control for activity of cycloheximide¤ keratinocyte lysates were prepared from cells treated with cycloheximide (+CHX) or left untreated (−CHX) and subsequently were maintained in normal growth medium (+Ca²⁺) or switched to low-calcium medium (−Ca²⁺). Fifteen micrograms of protein from each lane was resolved by SDS-PAGE, Western blotted, and immunoblotted with antibody to E-cadherin (C, lanes 1–4) or c-myc (C, lanes 5–8).