Supplemental file

Table S1. Oligo sequences shown in 5´ to 3´ orientation.

AH221 / ATGGCNGTNGCNTTYTAYATHCC
AH439_H3-F / CGAATTTCACGCAAAGCCACTG
AH440_H3-R / GCTCGTAAATCTGCTCCTTCAACC
AH600F / GCRAARAANCCRCAICCCAT
AH606F / GCNTTYTAYATHCCNGAYCARGC
AH639F / AGCGAGCATGAGACCAGCTGGGA
AH647F / CTTCACCACAACGAAATGGCCTTC
AH648R / GCGCTGTGTTGCCTTTCAATGGAGA
AH655R / TGCTGCTGGTGCTTCACATTTCGG
AH672 / AATCCGCGGATGGCCTTCGCTTACTA
AH673 / TATGGATCCTCAATGGAGAAATCTGG
AH675 / GCTCGCCAAGCCTATGTGCAGTATGAGCGC
AH676 / GCGCTCATACTGCACATAGGCTTGGCGAGC
AH677_rpr-F / CCCCGACCAGGCTACGCTATTG
AH678_rpr-R / AGTCGACATCTGCCCATGCTTTG
AH681_hid-F / AATCCGCGGATGGCTGTTGTCTT
AH683_hidIBM-F / AATCCGCGGATGGAGGCGGGCGCCGATGAC
AH687_hid-R / TAAGGATCCACTCATCGTGCCACAAGGAA
AH707_hidGH3-R / ATGCGGCCGCTCAGATGAAGGCTTTGAATTGCT
AH708_hid-F / AGGCAGTGGTGGCGGTGGCGAAGTA
AH709_hid-R / TGGGGTGGCTGTTGGAGGTGTTGTGT


Table S2. Statistical analysis. In addition to the t-test for each sample versus the control (indicated in Fig. 2, 3, and 4), the differences in the mean values among the specific treatment groups were calculated by one-way ANOVA using the Holm-Sidak method for pairwise multiple comparison procedures within SigmaPlot 11.

Fig. 2A. As-hid and As-rpr compared to their variants and homologs in AsE01

Comparison Diff of Means t P P<0,050

As-hid-IBM vs. As-hid 0,926 60,118 <0,001 Yes

As-hid-GH3 vs. As-hid 0,673 14,078 <0,001 Yes

Dm-hid vs. As-hid 0,266 11,044 <0,001 Yes

Dm-hidAla5 vs. As-hid 0,196 16,074 <0,001 Yes

Dm-hid vs. Dm-hidAla5 0,0700 4,411 0,012 Yes

As-rpr-IBM vs. As-rpr 0,728 16,026 <0,001 Yes

As-rpr-GH3 vs. As-rpr 0,652 11,881 <0,001 Yes

Dm-rpr vs. As-rpr 0,291 11,235 <0,001 Yes

Fig. 2B. As-hid and As-rpr compared to their variants and homologs in S2 cells

Comparison Diff of Means t P P<0,050

As-hid-IBM vs. As-hid 88,515 40,368 <0,001 Yes

As-hid-GH3 vs. As-hid 39,259 7,791 0,001 Yes

As-hid vs. Dm-hid 3,364 8,126 0,001 Yes

As-hid vs. Dm-hidAla5 4,010 10,846 <0,001 Yes

As-rpr-IBM vs. As-rpr 67,288 16,530 <0,001 Yes

As-rpr-GH3 vs. As-rpr 73,040 20,299 <0,001 Yes

As-rpr vs. Dm-rpr 20,987 13,835 <0,001 Yes

Fig. 3A. Co-operative function of hid and rpr in AsE01 cells

Comparison Diff of Means t P P<0,050

2x As-hid vs. As-hid +As-rpr 0,0457 3,558 0,024 Yes

2x As-rpr vs. As-hid +As-rpr 0,125 4,800 0,009 Yes

2x Dm-hid vs. Dm-hid + Dm-rpr - - 0,939 No

2x Dm-rpr vs. Dm-hid +Dm-rpr 0,103 7,842 0,001 Yes

Fig. 3B. Co-operative function of hid and rpr in S2 cells

Comparison Diff of Means t P P<0,050

2x As-hid vs. As-hid +As-rpr - - 0,127 No

2x As-rpr vs. As-hid +As-rpr 14,476 6,067 0,004 Yes

2x Dm-hid vs. Dm-hid +Dm-rpr 1,938 5,660 0,005 Yes

2x Dm-rpr vs. Dm-hid +Dm-rpr 2,416 6,957 0,002 Yes

Fig. 4A. As-hid:DIAP1 and As-rpr:DIAP1 in AsE01 cells

Comparison Diff of Means t P P<0,050

(As-hid : DIAP1)

1:0 vs. 1:1 0,0530 5,555 0,005 Yes

1:0 vs. 1:2 0,352 22,955 <0,001 Yes

1:0 vs. 1:4 0,700 34,271 <0,001 Yes

1:1 vs. 1:2 0,299 18,646 <0,001 Yes

1:1 vs. 1:4 0,647 30,872 <0,001 Yes

1:2 vs. 1:4 0,348 14,403 <0,001 Yes

(As-rpr : DIAP1)

1:0 vs. 1:1 - - 0,126 No

1:0 vs. 1:2 0,264 5,231 0,006 Yes

1:0 vs. 1:4 0,791 44,002 <0,001 Yes

1:1 vs. 1:2 - - 0,051 No

1:1 vs. 1:4 0,703 16,414 <0,001 Yes

1:2 vs. 1:4 0,527 10,944 <0,001 Yes

Fig. 4B. As-hid:DIAP1 and As-rpr:DIAP1 in S2 cells

Comparison Diff of Means t P P<0,050

(As-hid : DIAP1)

1:0 vs. 1:0.5 5,433 8,179 0,001 Yes

1:0 vs. 1:1 11,931 14,346 <0,001 Yes

1:0 vs. 1:2 47,845 22,563 <0,001 Yes

1:0.5 vs. 1:1 6,498 9,624 <0,001 Yes

1:0.5 vs. 1:2 42,412 20,547 <0,001 Yes

1:1 vs. 1:2 35,914 16,909 <0,001 Yes

(As-rpr : DIAP1)

1:0 vs. 1:0.5 - - 0,431 No

1:0 vs. 1:1 35,117 3,257 0,031 Yes

1:0 vs. 1:2 - - 0,056 No

1:0.5 vs. 1:1 - - 0,333 No

1:0.5 vs. 1:2 - - 0,460 No

1:1 vs. 1:2 - - 0,696 No

Fig. 5. Expression of As-hid and As-rpr in larvae and pupae

Comparison Diff of Means t P P<0,050

(As-hid)

E3L vs. 2L 0,198 10,486 <0,001 Yes

L3L vs. 2L 10,093 6,086 0,004 Yes

1d pupae vs. 2L 35,609 23,209 <0,001 Yes

8d pupae vs. 2L 6,637 18,970 <0,001 Yes

L3L vs. E3L 9,895 5,966 0,004 Yes

1d pupae vs. L3L 25,516 11,294 <0,001 Yes

1d pupae vs. 8d pupae 28,972 18,410 <0,001 Yes

(As-rpr)

E3L vs. 2L - - 0,860 No

L3L vs. 2L 11,294 13,932 <0,001 Yes

1d pupae vs. 2L 10,201 3,024 0,039 Yes

8d pupae vs. 2L 7,979 6,030 0,004 Yes

L3L vs. E3L 11,247 13,255 <0,001 Yes

1d pupae vs. L3L - - 0,768 No

1d pupae vs. 8d pupae - - 0,573 No


Fig. S1. ClustalW alignment of the full length Hid proteins in indicated species. Numbers on the right indicate the amino acid position in the protein.

Fig. S2. ClustalW alignment of the full length Rpr proteins in indicated species. Numbers on the right indicate the amino acid position in the protein.


As_Rpr MAFAYYIPDQATLLRRANLQEQQILRARQAYWRFVLTLIWDLLHQYVQYERQSMGRCRLPRFLH

DSC cccccccchhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhccccccccccc

MLRC ceeeeeccchhhhhhhhchhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhccccccccccccc

PHD cceeeccchhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhcccccccccc

Sec.Cons. cceeeccchhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhccccccccccc

Dm_Rpr MAVAFYIPDQATLLREAEQKEQQILRLRESQWRFLATVVLETLRQYTSCHPKTGRKSGKYRKPSQ

DSC ccccccccchhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhccccccccccccccccccc

MLRC ceeeeeccchhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhccccccccccccccccc

PHD ceeeeeccchhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhccccccccccccccccc

Sec.Cons. ceeeeeccchhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhccccccccccccccccc

Fig. S3. Predicted secondary structure of As_Rpr compared to Dm_Rpr. The consensus prediction server at PBIL (http://npsa-pbil.ibcp.fr/cgi-bin/npsa_automat.pl?page=/NPSA/npsa_seccons.html) was used to generate the shown secondary structures. Default combination methods (DSC, MLRC and PHD) were used to predict consensus (1). H – Alpha helix; C – Random coil; E - Extended strand.

1. Combet C, Blanchet C, Geourjon C, Deleage G. (2000) NPS@: network protein sequence analysis. Trends Biochem Sci 25:147-150.

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