Figure 4.

Subcellular localization of Vps28A and Vps28B and co-expression of Vps28A or Vps28B with Vps23A. BY-2 cells were co-transformed the indicated gene constructs and then cells were processed, imaged, and analyzed either by (A,B) epifluorescence microscopy or [(B) bottom row and (C)] CLSM as described in the legend of Figure 2. The yellow color in the corresponding merged images indicates co-localization between co-expressed proteins; white arrowheads also indicate obvious protein co-localization in the large, ring-like MVB-related structures. Note that the CLSM images presented in [(B) bottom row] are three different rotations (i.e., left ∼−15°, center, and right ∼+15°) obtained from the same 3D projection (full z-series) of immunostained Myc-Vps28A in an Myc-Vps28A and HA-Vps23A co-transformed cell; refer also to Figure S4 in Supplementary Material for the corresponding movie. The CLSM images of the triple-transformed cell in (C) are single z-sections. Bar = 10 μm.