PEGylation of olive pollen OBs and localization of caleosin by fluorescence microscopy. (A) Localization of caleosin on PEGylated (+) and non-PEGylated (-) OBs from pollen using a FL anti-Clo3 caleosin Ab, followed by a secondary Ab conjugated with DyLight 549. After PEGylation, the accessibility of the FL anti-Clo3 Ab was hampered and no fluorescence was observed. (B) Localization of caleosin as above but using an N-terminal (αN) anti-Clo3 caleosin Ab. After PEGylation, the primary Ab was able to bind to caleosin.