1. GFP silencing reverted cells X GFP silenced cells Vectors employed GFP expressing cells Sf21 cell Figure 1A Experimental line-1 Experimental line-2 Control line

2. a. b. c. f. 1 2 3 1 2 i ii d. 1 2 3 21 nt 1 M 2 14 kDa gfp shRNA (49 bp) EcoRI 19 bp9 bp19 bp 23 nt PCR RT-PCR  -actin loading control Figure 1B 0 4000 2000 6000 1 2 3 4 e. i ii iii mRNA Total RNA loading control KpnI gfp shRNA Forward primer gfp shRNA Reverse primer

3. Figure 2 10 0 Counts FLH-1 10 2 10 1 10 3 10 4 0 120 40 80 160 200 A B Visible Light UV Light i ii iii

4. 800 1600 2400 0 GFP expressing line FHVB2- C17 line GFP silencing line FHVB2 expressing line Relative intensity of siRNA Total RNA loading control siRNA Figure 3 (ii) (iii) 1 2 3 4 (i)

5. Figure 4 (i) (ii) (iii) 13 14 15 16 17 18 Catalase Aldolase B A C 1 2 3 M kDa 160 110 73 D 10 11 12 13 14 15 16 17 18 19 20 21 22 23 E 23mer M 1 2 21mer 0 200 400 600 800 1000 1200 1400 1600 mAU 0.05.010.015.020.0ml 14 16 15

6. 0.0 5.010.015.020.0 0 200 400 600 800 1000 1200 1400 1600 mAU A 25.0ml B i ii 10 11 12 13 14 15 16 17 18 19 20 21 22 23 +ve C 15 14 16 14 15 16 Pool W. Probe  - FHVB2  -DCR-1 PAZ Figure 5

7. Figure 6 A SD (Leu - Trp - His - ) SD (Leu - Trp - ) (v) (vi) YPD (ii)(iii) SD (Trp - )SD (Leu - ) (iv) (i) Scheme pGADT7+ pGBKT7 pGBKT7 pGADT7 AH109 AD (REP) +BD (REP) AD(ENDO) BD(FHV B2) pGBKT7 FHVB2 AD(PAZ) BD(FHV B2) B (i) (ii) M 1 2 3 1 2 3 4 M 5 6 116 66 45 35 25 18 (iii) M 1 2 3 INPUTELUTES FLOW THROUGH FT I EL

8. Figure 7 M 1 2 3 4 5 6 7 INPUT B GST-FHVB2 MBP-DCR-2-PAZ MBP EL FT 118 81 52 36 kDa A AH109 pGAD + pGBD pGAD AD(DCR2PAZ) +BD (FHVB2) pGBD (FHVB2) AD(Rep)+ BD(Rep) pGBD AD (DCR2 PAZ)+ pGBD (i) Scheme(ii) SD (Leu - Trp - )(iii) SD (Leu - Trp - His - )

9. Full Length FHVB2 (107aa) FHVB2 – C17 (90aa) N N C C Figure 8 A B C Sf21 cell line GFP expressing GFP silenced FHVB2- GFP reverted FHVB2- C17 transfected 0 0.2 0.4 0.6 0.8 1.0 1.2 GFP GFP silenced GFP reverted FHVB2- C17 Relative level of gfp mRNA % % % % % ( )

10. Figure 9 1 2 M 3 4 5 M 6 7 B INPUT ELUTE F T (i) (i) Scheme (ii) SD (Leu - Trp - His - ) A (iv) SD (Leu - Trp - His - )(iii) Scheme AD+BD BD AD BD(FHV-B2) AH109 AD(REP) +BD(REP) AD- DCR1 (PAZ)+ BD(FHVB2-  C17) AD(PAZ) DCR1+BD (FHVB2) AD+BD BD AD BD(FHV-B2) AH109 AD(REP) +BD(REP) AD- DCR2 (PAZ)+ BD(FHVB2-  C17) AD(PAZ) DCR1+BD (FHVB2) ELUTE FT INPUT kDa M 1 2 3 (ii) 36 52 81 118

11. PAZ 2249 amino acids N-terminal C-terminal 544aa 606aa825aa DUF 283 920aa HELICASE-C 510aa1096 aa1218 aa1706 aa RNase III 1919 aa dsRNA bd 2241 2175aa aa MBP-PAZ MBP-ENDONUCLEASE dm Dicer-1 dm Dicer-2 PAZ 1715 amino acids N-terminal C-terminal 535aa 554aa659aa HELICASES 1aa838aa945aa1187aa RNase III 1642aa dsRNA bm 1710aa 1646aa MBP-PAZ dsRNA bd A. B. C. Supplementary Figure 1 dm Dicer-1-PAZ dm Dicer-2-PAZ dm Dicer-1-PAZ dm Dicer-2-PAZ dm Dicer-1-PAZ dm Dicer-2-PAZ