Figure 3.

Exocyst depletion induces the accumulation of CD63-positive MVBs

(A–D) CD63 expression in HN4 cells after the cell were transfected with siCtrl or with Exo70, Sec3, or Sec10 siRNA. (A) CD63 immunofluorescence in HN4 cells accumulates after transfection with Exo70 small interfering (si)RNA, Sec3 siRNA, or Sec10 siRNA versus with scrambled siRNA. Scale bar, 50 μm. (B) Quantification analysis of fluorescence intensity of CD63. (C) Representative western blotting images of CD63 expression. (D) Quantification of CD63. β-Tubulin was used for normalization. ∗p < 0.05, ∗∗p < 0.01 versus siCtrl by t test. (E–G) The number of MVBs and ILVs after transfection with scrambled siRNA, Exo70 siRNA, Sec3 siRNA, or Sec10 siRNA. (E) Representative electron microscopy images of HN4 cells after transfection with scrambled siRNA, Exo70 siRNA, Sec3 siRNA, or Sec10 siRNA. Scale bar, 1 μm (F) The number of MVBs per 100 μm². (G) The number of ILVs per MVB. ∗p < 0.05, ∗∗p < 0.01, ∗∗p < 0.01 versus siCtrl by t test.