Figure 3
The TRAPPC10 and TRAPPC11 ASH domains target to the centrosome and affect microtubule organization. RPE cells expressing Myc-TRAPPC10-ASH or Myc-TRAPPC11-ASH were serum-starved for 24 h, fixed with methanol (A) or PFA (B, C) and stained with rabbit monoclonal antibody against Myc and mouse monoclonal antibody against α-tubulin (tub), acetylated tubulin (Ac tub) or p150Glued, as indicated. DNA was stained with DAPI. Note that the Myc-ASH fusion proteins localize to the centrosome (arrowheads) and basal body (asterisk), and cause bundling of microtubules (open arrow). An example of a primary cilium, stained with Ac tub antibody, is indicated with a closed arrow. Punctate staining near the nucleus can be seen in some cells expressing Myc-ASH fusion proteins (green staining in panel A). Insets show enlargement of the centrosome area.