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S2 Supporting information: RT-qPCR experiment

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1 S2 Supporting information: RT-qPCR experiment
Housekeeping gene description (slide 2) Explanatory slide (slide 8) Detailed validating tests (slide10)

2 1. Housekeeping gene description

3 RNAseq experiment: no sex-bias of expression
Smp_ (Wang et al. 2013) Sm-fad oxidoreductase, FAD dependent oxidoreductase domain containing protein RNAseq experiment: no sex-bias of expression Expression (rpkm) Males Females Cercariae Adults S#1 S#2 S#3 (Rpkm) Cerc. S#1 S#2 S#3 Ad. Male Female

4 RNAseq experiment: no sex-bias of expression
Smp_ (Wang et al. 2013) Sm-RPL35, subunit of the oligosaccharyltransferase RNAseq experiment: no sex-bias of expression Expression (rpkm) Males Females Cercariae Adults S#1 S#2 S#3 (Rpkm) Cerc. S#1 S#2 S#3 Ad. Male Female

5 RNAseq experiment: no sex-bias of expression
Smp_ (Wang et al. 2013) Sm-arp 10, actin protein 10 RNAseq experiment: no sex-bias of expression Expression (rpkm) Males Females Cercariae Adults S#1 S#2 S#3 (Rpkm) Cerc. S#1 S#2 S#3 Ad. Male Female

6 RNAseq experiment: no sex-bias of expression
Smp_ (Wang et al. 2013) Aspartyl-tRNA synthetase NOT SELECTED BECAUSE OF BAD AMPLIFICATION EFFICIENCY RNAseq experiment: no sex-bias of expression Expression (rpkm) Males Females Cercariae Adults S#1 S#2 S#3 (Rpkm) Cerc. S#1 S#2 S#3 Ad. Male Female

7 RNAseq experiment: sex-bias of expression!
Smp_090120 Alpha tubuline NOT SELECTED BECAUSE OF SEX-BIAS OF EXPRESSION IN RNAseq RNAseq experiment: sex-bias of expression! Expression (rpkm) * Males Females * Cercariae Adults S#1 S#2 S#3 (Rpkm) Cerc. S#1 S#2 S#3 Ad. Male Female

8 2. Explanatory slide

9 No significative bias in RNAseq
Genome v5.02 Gene ID Cufflinks ID Functional annotation  Functional category A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression Significative female-bias by RNAseq (P<0,05; DEseq) * ♀ ♀ No significative bias in RNAseq ♂ ♂ 3 developmental stages Sexual dimorphism 5 developmental stages Sexual dimorphism Spearman Rank test taking in consideration the three stages (Rank correlation between the two experiments) Coefficient R P value Males Females

10 3. Detailed tests

11 Significantly correlated genes between RNAseq & qPCR: 30

12 Smp_065610 XLOC_005850 Pyruvate kinase  Cell metabolism
A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * * * 3 developmental stages 5 developmental stages Coefficient R P value 0,9046 <0,0001 Males Females

13 DNA replication licensing factor mcm6
Smp_ XLOC_031213 DNA replication licensing factor mcm6  DNA modifications A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,9612 <0,0001 Males Females

14 DNA repair protein rad51 homolog 1
Smp_ XLOC_000295 DNA repair protein rad51 homolog 1  DNA modifications A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,9329 <0,0001 Males Females

15 Pyruvate dehydrogenase
Smp_ XLOC_025837 Pyruvate dehydrogenase  Cell metabolism A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * * 3 developmental stages 5 developmental stages Coefficient R P value 0,9188 <0,0001 Males Females

16 Invadolysin (m08 family)
Smp_ XLOC_015382 Invadolysin (m08 family)  Host-Pathogen interactions Proteases and related proteins A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * * 3 developmental stages 5 developmental stages Coefficient R P value 0,9912 <0,0001 Males Females

17 Long-chain-fatty-acid ligase
Smp_ XLOC_022547 Long-chain-fatty-acid ligase  Cell metabolism A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * * * 3 developmental stages 5 developmental stages Coefficient R P value 0,9753 <0,0001 Males Females

18 Lysosomal pro-xaa carboxypeptidase (s28 family)
Smp_ XLOC_013222 Lysosomal pro-xaa carboxypeptidase (s28 family)  Vesicular compartment Reads count (rpkm) Relative expression * * 3 developmental stages 5 developmental stages Coefficient R P value 0,9329 <0,0001 Males Females

19 Smp_063040 XLOC_028567 Actin-like protein 6a
 Cytoskeletal, structural & motor proteins Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,9046 <0,0001 Males Females

20 Anti-silencing protein (ASF1 like histone chaperone)
Smp_ XLOC_025447 Anti-silencing protein (ASF1 like histone chaperone)  Chromatin structure A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,8764 0,0002 Males Females

21 Poly(adp-ribose) glycohydrolase (PARG)
Smp_ XLOC_022651 Poly(adp-ribose) glycohydrolase (PARG)  Chromatin structure A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * * 3 developmental stages 5 developmental stages Coefficient R P value 0,8764 0,0002 Males Females

22 Structural maintenance of chromosomes protein 2 (SMC2)
Smp_ XLOC_022066 Structural maintenance of chromosomes protein 2 (SMC2)  Chromatin structure A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,8198 0,0011 Males Females

23 Family c50 non-peptidase homologue (c50 family)
Smp_ XLOC_026909 Family c50 non-peptidase homologue (c50 family)  Undefined Functions A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,8057 0,0016 Males Females

24 Glucose-6-phosphate isomerase
Smp_ XLOC_024997 Glucose-6-phosphate isomerase  Cell metabolism Reads count (rpkm) Relative expression * * 3 developmental stages 5 developmental stages Coefficient R P value 0,8057 0,0016 Males Females

25 Porphobilinogen deaminase
Smp_ XLOC_022698 Porphobilinogen deaminase  Host-Pathogen interactions Exploitation of host microenvironment A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,8057 0,0016 Males Females

26 Smp_031720 XLOC_021276 Histone h2a  Chromatin structure
A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,7916 0,0022 Males Females

27 Smp_040020 XLOC_018236 Histone h3  Chromatin structure * *
Reads count (rpkm) Relative expression * * 3 developmental stages 5 developmental stages Coefficient R P value 0,7633 0,0039 Males Females

28 Prospero homeobox protein 1
Smp_ XLOC_028377 Prospero homeobox protein 1  Development A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,7633 0,0039 Males Females

29 Camp-dependent protein kinase catalytic subunit alpha
Smp_ XLOC_026035 Camp-dependent protein kinase catalytic subunit alpha  Signalling Pathways Reads count (rpkm) Relative expression * * 3 developmental stages 5 developmental stages Coefficient R P value 0,7492 0,005 Males Females

30 Pur-alpha-like protein
Unknown Smp XLOC_012827 Pur-alpha-like protein  Transcription associated proteins Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,735 0,0065 Males Females

31 Smp_005210 XLOC_001572 Histone deacetylase 1  Chromatin structure *
Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,6785 0,0153 Males Females

32 S-phase kinase-associated protein 1
Smp_ XLOC_012583 S-phase kinase-associated protein 1  Cell cycle A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,6785 0,0153 Males Females

33 Leishmanolysin-like peptidase
Smp_ XLOC_016494 Leishmanolysin-like peptidase  Host-Pathogen interactions Proteases and related proteins A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,6643 0,0184 Males Females

34 Caax prenyl protease 1 homolog
Smp_ XLOC_010022 Caax prenyl protease 1 homolog  Host-Pathogen interactions Proteases and related proteins A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,6502 0,0221 Males Females

35 Leucyl aminopeptidase
Smp_ XLOC_012112 Leucyl aminopeptidase  Cell metabolism A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,6361 0,0262 Males Females

36 Nucleosome assembly protein (NAP)
Smp_ XLOC_012229 Nucleosome assembly protein (NAP)  Chromatin structure A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,6219 0,0308 Males Females

37 Histone-lysine n-methyltransferase
Smp_ XLOC_017321 Histone-lysine n-methyltransferase  Chromatin structure A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * * 3 developmental stages 5 developmental stages Coefficient R P value 0,6219 0,03308 Males Females

38 Mixed-lineage leukemia
Smp_ XLOC_002973 Mixed-lineage leukemia  Chromatin structure A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,5937 0,0418 Males Females

39 Smp_026560 XLOC_024515 Calmodulin  Signalling Pathways *
Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,5513 0,0432 Males Females

40 Paired box protein pax-6
Smp_ XLOC_017450 Paired box protein pax-6  Development A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,5795 0,0483 Males Females

41 Protein kinase c alpha type
Smp_ XLOC_031864 Protein kinase c alpha type  Signaling pathways A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,5795 0,0483 Males Females

42 Non significantly correlated genes between RNAseq & qPCR: 10

43 Organic solute transporter
Smp_ XLOC_020282 Organic solute transporter  Channels and transporters Reads count (rpkm) Relative expression * * 3 developmental stages 5 developmental stages Coefficient R P value 0,5654 0,0554 Males Females

44 Serine threonine protein kinase
Smp_ XLOC_005397 Serine threonine protein kinase  Signalling Pathways Reads count (rpkm) Relative expression * * 3 developmental stages 5 developmental stages Coefficient R P value 0,523 0,081 Males Females

45 Smp_089860 XLOC_010265 Histone H3.2  Chromatin structure *
Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,4806 0,1138 Males Females

46 Syntrophin-like isoform e
Smp_ XLOC_025116 Syntrophin-like isoform e  Undefined Functions Protein-Protein interactions A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,3958 0,2028 Males Females

47 26s proteasome regulatory subunit n11
Smp_ XLOC_015958 26s proteasome regulatory subunit n11  Protein processing Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,3958 0,2028 Males Females

48 Voltage-gated calcium chanel
Smp_ XLOC_000749 Voltage-gated calcium chanel  Transporters and channels A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,2544 0,4249 Males Females

49 Ubiquitin-conjugating enzyme e2
Smp_ XLOC_031143 Ubiquitin-conjugating enzyme e2  Signalling Pathways Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,1696 0,5982 Males Females

50 T-box transcription factor tbx2
Smp_145230 T-box transcription factor tbx2  Development A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,07068 0,8272 Males Females

51 Probable histone-binding protein caf1-like
Smp_ XLOC_022332 Probable histone-binding protein caf1-like  Chromatin structure Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,04241 0,8959 Males Females

52 Smp_085290 XLOC_007127 Tropomyosin  Cytoskeleton-related proteins
A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value 0,01414 0,9652 Males Females

53 Inversely correlated genes between RNAseq & qPCR: 3

54 Nucleosome assembly protein (NAP)
Smp_ XLOC_029007 Nucleosome assembly protein (NAP)  Chromatin structure A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value -0,3958 0,2028 Males Females

55 Smp_061510 XLOC_018289 Nardilysin (m16 family)
 Host-Pathogen interactions Proteases and related proteins A. RNA-seq experiment B. qPCR experiment Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value -0,04241 0,8959 Males Females

56 Smp_166110 XLOC_014666 Ebna2 binding protein p100
 Transcription factor Reads count (rpkm) Relative expression * 3 developmental stages 5 developmental stages Coefficient R P value -0,04241 0,8959 Males Females

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