Figure 4.

LlLIM1 overexpression caused the mislocalization of signal molecules involved in the PLC signaling pathway in the clear zone of elongating pollen tubes. The images show the typical (A) and missed (B and C) subcellular localization of the fluorescent DAG marker, CYS1:GFP, in CFP-co-overexpressed (A) or LlLIM1-co-overexpressed (B and C) pollen tubes stained with FM4-64 to show the distribution of secreted/endocytic vesicles. B and C represent pollen tubes showing slow and stopped growth, respectively. In all experiments, pollen grains were cobombarded with 2.5 μg of expressing plasmid of the indicated florescent markers and 5 μg of expressing plasmid of CFP or LlLIM1, cultured in germination medium for 6 h, and then confocal images were recorded with the proper fluorescent channels. Images were obtained from the central section of pollen tubes lying flat on the cover-slide surface and represent typical examples from at least 10 similar images collected from at least three independent experiments.