7.2@ Cytoskeletal
inhibitors
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)
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 |