Hello Sungho Jeong

Yes, you should add cytochalasin D as part of the protocol.

Actin-containing cytoskeletal network plays an important role in phagocytosis and endocytosis. Cytochalasin D is used to assess the role of the actin cytoskeleton in different biological processes including endocytosis. Cytochalasin D is a cell-permeable and potent inhibitor of actin polymerization. It disrupts actin microfilament. It is believed to bind to F-actin polymer and prevent polymerization of actin monomers.

Cytochalasin D treatment severely disrupts network organization, increases the number of actin filament ends, and leads to the formation of filamentous aggregates or foci composed mainly of actin filaments. So, to assess the function of a biological process such as endocytic capacity of dendritic cells in this case, the use of an inhibitor like Cytochalasin D becomes essential.

I hope this helps.

Best.

Thanks Malcolm Nobre !

That really helped me understand the purpose of using that reagent.

Many thanks

Sungho

Hi Singho,

The answer of Malcolm Nobre is the really correct and as u said that helped you to get the result also.

I juss wantd to add more, immature DC are at their endocytic stage and they are prone for endocytosis process to uptake. As DC is immune cells and APC so they follow the phagocytosis as well as macropinocytosis also. Cytochalasin D is a selective inhibitor of both the endocytosis process and mechanisms behind that beautifully described above.

As there are many other pathways of endocytosis such as clathrin, caveolae dependent and non dependent. If u use some other inhibitors aslo such as chlorpromazine, dynasore, statins you can represent ur data more precisely.

All the best.