Figure 3
Expression analysis of lily PEPCs in male tissues and developing pollen. (A) Expression of LlBTPC and LlPTPC in anther tissues. Proteins (10 μg) from the pollen and anther wall of 14 cm flower buds were treated with anti-AtBTPC (upper) or anti-AtPTPC (middle) antibody. Non-specific reactive bands were used as loading control (bottom). The upper bands detected with anti-AtPTPC antibody (asterisk) represent Ub-LlPTPC. (B) Expression patterns of LlBTPC and LlPTPC in developing microspores and pollen. Top diagram illustrates lily pollen development, each stage of which corresponds to the length of the flower bud (cm). Protein extracts from lily pollen (10 μg) were subjected to immunoblot analysis with anti-AtBTPC (upper) or anti-AtPTPC (middle) antibody. The upper bands detected with anti-AtPTPC antibody (asterisk) represent Ub-LlPTPC. Non-specific reactive bands were used as loading control (bottom). PMI, pollen mitosis I; MS, microspore; EB, early bicellular pollen; MB, mid bicellular pollen; LB, late bicellular pollen; GC, generative cell; VC, vegetative cell; PA, post anthesis. (C) Nondenaturing PAGE analysis of microspore and late bicellular pollen proteins (7 μg). The numbers in each panel represent the length of the flower bud (cm) containing the microspores (3) or late bicellular pollen (14). In-gel staining for PEPC activity (left), and immunoblot analysis with anti-AtPTPC (middle) or anti-AtBTPC (right) antibody were performed.