Which of the following methods is most suitable to determine

Which of the following methods is most suitable to determine if the actin cytoskeleton is dynamic and constantly undergoes assembly and disassembly?

a.Immunofluorescence microscopy with a primary antibody to G-actin (raised in mouse) and a secondary anti-mouse antibody (raised in goats) coupled to a green fluorophore.

b. Live imaging of cells expressing GFP-tagged actin

c. FRAP analysis in cells expressing GFP-tagged actin

d. Electron microscopy

Solution

Answer: b. Live imaging of cells expressing GFP-tagged actin

Reason:

During cell division at the time of wound healing fibroblasts are going to protrude through -actin polymerization & depolymerization can be observed using live imaging of cells expressing GFP-tagged actin & F-actin marker, and using different genetically combining fluorescent proteins with different actin-binding domains. The protrusions in the cells are going to produce a lamellopodium for cell migration & this process is mediated by “calcium independent myosinII kinase light chain in these cells (Non-muscle cells) and these are detected using imaging of live cells of GFP-labeled. Retraction movements of cell during cell migration is mediated by the “the adhesions” & the movement of cells is mediated by Rho, Rac, and Cdc42 proteins meticulously.