7.2@ Cytoskeletal inhibitors

Yoshikatsu Sato

 

Preparation

Microtubule depolymerizing drugs

Oryzalin@@@@@@@@@ 2 mM in DMSO (stock) à 1-10 µM (final)

Cremart@@@@@@@@@ 2 mM in DMSO (stock) à 10-20 µM (final)

 

 

Actin filament depolymerizing drugs

Cytochalasin B@@@@ 2 mM in DMSO (stock) -à 10 µM (final)

Latrunculin B@@@@@ 2 mM in DMSO (stock) -à 10 µM (final)@ (Sigma, L-5288)

 

 

Procedure

1. Incubate protonemata between two layers of an agar-gelatin film on a coverslip and culture under unilateral red light (0.5 Wm-2) for 1-2 weeks in the liquid BCDAT medium (See 2.5).

2. Mix the stock solution (above) with BCDAT medium according to a table below, and replace the medium with the inhibitor-containing medium.@

 

 

DMSO control

10 µM Oryzalin

0.1 mM Cytochalasin B

Oryzalin + Cytochalasin B

BCDAT medium

3960 µl

3960 µl

3960 µl

3960 µl

2 mM Oryzalin

-

20µl

-

20µl

20 mM Cytochalasin B@@@@@ @@@@@@@

-

-

20µl

20µl

DMSO (1%)

40 µl

20µl

20µl

 

Total

4000 µl

4000 µl

4000 µl

4000 µl