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. 2017 Feb 21;14(2):213. doi: 10.3390/ijerph14020213

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© 2017 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Western blot of AeCA9 in each life stage, female posterior midgut, and female Malpighian tubules. (A) In this western blot, the same life stages, sexes, and tissues were analyzed as in Figure 7. 6 μg of total protein was loaded for the life stages (4th instar larvae, Male pupae, Female pupae, Male adult, Female adult) and 40 μg of total protein was loaded for the female posterior midugt and female Malpighian tubules. AeCA9 was detected in all the life stage and tissue samples analyzed except the MTs (top row). There is a higher molecular weight band in the female adult sample, but that could be due to over-development; (B) This is an alignment showing the antigenic sequence used to develop the antisera targeting AeCA9 in (A) and all subsequent immunohistochemical analyses. The antisera was generated in chicken to target Anopheles gambiae CA9, and it has suitable cross-reactivity with Aedes aegypti as shown. Take note that there is only 1 amino acid difference in 19 between the antigenic sites of AgCA9 and AeCA9.