Table 3. Evaluation of the assay system 3: ebony as the marker gene.

1st RNAi	2nd RNAi		Initial # of larvae	# survived	# of 2nd instar	ebony	WT	suppression
250 bp Ago2 (5 ug/mL)	250 bp ebony (5 ug/mL)	co-feeding	30	24	18	13	5	27%
250 bp Dcr2 (5 ug/mL)	250 bp ebony (5 ug/mL)	co-feeding	28	26	25	19	6	24%
250 bp KA (5 ug/mL)	250 bp ebony (5 ug/mL)	co-feeding	27	24	19	13	6	31%
250 bp Ago2 (5 ug/mL)	250 bp ebony (5 ug/mL)	sequential	30	20	16	13	3	18%
250 bp Dcr2 (5 ugmL)	250 bp ebony (5 ug/mL)	sequential	28	21	20	19	1	5%
250 bp KA (5 ug/mL)	250 bp ebony (5 ug/mL)	sequential	27	14	13	13	0	0%

Evaluation of the assay system with ebony as the marker gene instead of lac2. The second instar larvae were categorized into the ebony phenotype (i.e. the second RNAi worked) and the WT phenotype (i.e. the second RNAi is suppressed). The suppression of the second RNAi phenotype by the initial RNAi was evaluated as the proportion of the WT larvae in the survived second instar larvae (Suppression).