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