Figure 3.
High magnification CLSM indicates that DLC-AS–containing RPP can form a complex with LC8 that interacts with the MT cytoskeleton in live cells. Cells were transfected to express the indicated proteins and analyzed by CLSM (100×) without (top panel) or with (bottom panel) pretreatment with the MT-stabilizing agent, taxol. GFP-RPP174-297 showed no evidence of association with MT cytoskeleton when expressed alone (left panel) or coexpressed with dsRed-LC8 (right panel), and no effect of taxol treatment was apparent. At this magnification, GFP-RPP139-297 did not show association with cytoskeletal structures when expressed alone. When GFP-RPP139-297 was coexpressed with dsRed-LC8, a proportion of the transfected proteins colocalized at a perinuclear region and on filamentous structures that converged at the perinuclear region (see yellow colorations in merged image). Treatment with taxol did not affect the appearance of cells expressing GFP-RPP139-297 alone, but in cells coexpressing GFP-RPP139-297 and dsRed-LC8, the filamentous structures on which the transfected proteins were colocalized became more defined.