Table 9. Transfection to detect biphasic effect of arrestin-3 on ASK1-stimulated JNK phosphorylation (Fig. 6A,B).

pcDNA3-Arrestin-3	0 μg	0 μg	0.1 μg	0.25 μg	0.5 μg	1.0 μg	2.0 μg	4.0 μg
pcDNA3-HA-ASK1	0 μg	0.5 μg	0.5 μg	0.5 μg	0.5 μg	0.5 μg	0.5 μg	0.5 μg
pcDNA3	4.5 μg	4.0 μg	3.9 μg	3.75 μg	3.5 μg	3.0 μg	2.0 μg	0 μg
FuGENE HD	13.5μl	13.5μl	13.5μl	13.5μl	13.5μl	13.5μl	13.5μl	13.5μl
Opti-MEM	125μl	125μl	125μl	125μl	125μl	125μl	125μl	125μl

^*Note: These DNA amounts were calculated for 6-well culture plate (equivalent to 35 mm dish). If different size plates are used, the amount of total DNA should be changed proportionally to plate area.