Fig. 6.

Validation of RNAi-mediated silencing of AamS6 by RT-PCR and western blot analyses. At 48 h post attachment three ticks per treatment as indicated were manually detached and processed individually for total RNA and protein extraction as detailed in the Materials and methods. (A) Total RNA was subjected to two-step semi-quantitative RT-PCR to amplify the C-terminal transcript that is unique to AamS6, the N-terminal transcript that is conserved between AamS6 and AamS17, and the tick actin fragment (sample load control). (B) Total protein extracts from the same animals that were used for total RNA extraction in A were subjected to western blot analysis using antibodies to rAamS6.