Table 2.

Summary of RNAi screen candidates

Gene KD	N	Developing Oocysts						Melanized Ookinetes
		Md	Load*	Fd	P†	Pr (%)‡	χ2§	Load*	Fd	P†	Pr (%)‡	χ2§
Genes affecting oocyst load and prevalence of infection
AGAP004016¶	35	2	14 ± 3.9	0.4	0.004	63	1e−4	11 ± 0.7	9.4	0.064	42	0.007
LacZ		16	35 ± 7.1			86		1 ± 5.3			20
AGAP006914¶	47	5	27 ± 6.4	0.6	0.041	74	0.009	6 ± 2.8	1.4	0.307	45	0.009
LacZ		19	41 ± 7.9			83		5 ± 2.1			28
AGAP010325¶	41	13	28 ± 5.5	2.2	0.017	85	0.008	1 ± 0.4	0.3	0.212	20	0.048
LacZ		6	13 ± 3.5			66		3 ± 1.4			34
EST08518¶	47	2	13 ± 3.9	0.4	0.002	57	4e−4	13 ± 10	8.6	0.641	30	0.500
LacZ		19	36 ± 7.5			79		2 ± 0.6			26
Genes affecting oocyst load only
AGAP009642	54	24	49 ± 8.4	1.8	0.017	89	0.465	4 ± 1.4	2.2	0.341	32	0.233
LacZ		11	28 ± 6.6			86		2 ± 0.7			25
AGAP005761	45	15	43 ± 10	2.3	0.047	82	0.177	8 ± 4.9	2.9	0.540	22	0.197
LacZ		6	19 ± 5.2			77		3 ± 1.3			31
Genes affecting ookinete melanization load and prevalence of ookinete melanization
AGAP003304¶	52	11	40 ± 8.2	1.0	0.252	83	0.192	11 ± 3.7	2.5	0.021	54	3e−4
LacZ		17	39 ± 7.4			88		4 ± 1.8			31
AGAP012000¶	55	17	38 ± 6.9	0.9	0.441	82	0.715	17 ± 6.9	5.8	0.021	45	2e−6
LacZ		21	41 ± 7.4			84		3 ± 1.5			20
AGAP001508¶	42	7	30 ± 9.6	0.8	0.273	69	1.0	9 ± 5.1	1.5	0.040	77	7e−8
LacZ		16	38 ± 10			69		6 ± 4.7			17
Genes affecting prevalence of ookinete melanization
AGAP007314¶	32	2	14 ± 3.7	0.8	0.460	62	0.710	17 ± 8.6	3.2	0.118	53	0.008
LacZ		2	18 ± 8.1			66		5 ± 2.3			31
AGAP001242¶	48	30	57 ± 9.4	1.7	0.157	77	0.220	6 ± 3.2	6.7	0.144	31	0.007
LacZ		20	34 ± 7.2			85		1 ± 0.5			16
AGAP009380¶	57	6	13 ± 3.2	1.2	0.674	70	0.547	2 ± 0.8	0.2	0.099	17	0.011
LacZ		7	11 ± 1.9			66		6 ± 2.3			33

All significant P values (<0.05) and chi-square test values are highlighted in bold. KD, knocked down; N, number of mosquitoes tested per KD; Md, median; Fd, fold difference; Pr, prevalence.

*Load represents the arithmetic mean (+/− one Standard Error of the Mean) of parasite numbers, including uninfected guts, in three biological repeats.

^†Mann–Whitney U test on oocyst or melanized ookinete load.

^‡Percent sign refers to the percentage of infected mosquitoes.

^§Chi-square test on oocyst or melanized ookinete prevalence.

^¶All gene KDs with an effect on prevalence of infection and/or melanization were further analyzed to determine if parasite load is affected once prevalence effect is eliminated from the analysis by removing uninfected midguts from the dataset. Two gene KDs showed an additional effect of infection intensity: AGAP004016 (N = 22, dsGOI: 23 ± 5.5, dsLacZ: 61 ± 17, P = 0.02) and ENSANGESTG00000008518 (N = 27, dsGOI: 20 ± 6.2, dsLacZ: 50 ± 12; P = 0.007).