- Visualization of Actin Organization and Quantification in Fixed Arabidopsis Pollen Grains and Tubes.
Visualization of Actin Organization and Quantification in Fixed Arabidopsis Pollen Grains and Tubes.
Bio-protocol (2020-01-05)
Xiaolu Qu, Qiannan Wang, Haiyan Wang, Shanjin Huang
PMID33654717
ABSTRACT
Although it is widely accepted that actin plays an important role in regulating pollen germination and pollen tube growth, how actin exactly performs functions remains incompletely understood. As the function of actin is dictated by its spatial organization, it is the key to reveal how exactly actin distributes in space in pollen cells. Here we describe the protocol of revealing and quantifying the spatial organization of actin using fluorescent phalloidin-staining in fixed Arabidopsis pollen grains and pollen tubes. We also introduce the method of assessing the stability and/or turnover rate of actin filaments in pollen cells using the treatment of latrunculin B.
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Calcium chloride dihydrate, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%
Sigma-Aldrich
Latrunculin B, Latrunculia magnifica, Latrunculin B, CAS 76343-94-7, is a unique marine toxin that inhibits actin polymerization and disrupts microfilament organization. It is 10 to 100-fold more potent than cytochalasins.
Sigma-Aldrich
Dimethyl sulfoxide, sterile-filtered, BioPerformance Certified, meets EP, USP testing specifications, suitable for hybridoma
Sigma-Aldrich
Boric acid, BioReagent, Molecular Biology, suitable for cell culture, suitable for plant cell culture, ≥99.5%