Table 1

Primer for PCR"

Table 2

Sequence"

Fig. 1

Amplication of 5′UTR and 3′UTR DNA fragment by PCR M. 2000 DNA marker; lanes 1 and 4. 5′UTR-ds-T7; lanes 2 and 5. 5′UTR-s-T7; lanes 3 and 6. 5′ UTR-a-T7; lane 7. 3′UTR-△polyU; lane 8. 3′UTR-s-T7; lane 9. 3′UTR-a-T7"

Fig. 2

In vitro transcription bands were biotin-labeled for the 5′UTR and 3′UTR M. 2000 DNA marker; lane 1. DNA-5′UTR; lane 2. RNA-bio-ss(+) 5′UTR; lane 3. RNA-bio-ss(-) 5′UTR; lane 4. RNA-bio-ss(+) 3′UTR; lane 5. RNA-bio-ss(-)3′UTR; lane 6. RNA-bio-ss(-)SL1; lane 7. RNA-bio-ss (-) SL2; lane 8. 3′UTR-△polyU"

Fig. 3

Silver-stained gel images of 3′UTR RNA-interacting proteins M. 180 ku protein marker; lane 1. ss(-)5′UTR; lane 2. ss(+)5′UTR; lane 3. ss(-)3′UTR; lane 4. ss(+)3′UTR; lane 5. Negative"

Fig. 4

Venn diagram analysis of mass spectrometry data for ss(-)3′UTR and Negative"

Table 3

Mass spectrometry protein information"

Fig. 5

GO and KEGG analysis of 106 proteins interacting with the 3′UTR minus strand"

Fig. 6

Protein-protein interaction network"

Fig. 7

Verify Ddx18 protein and FMDV 3′UTR A. Dual enzyme digestion to identify pCAGGS-HA-Ddx18 eukaryotic expression vector (M. DNA marker; Lane 1.pCAGGS-HA-Ddx18); B. Western blot verification of the expression of Ddx18 protein (M.180 ku protein marker; Lane 1. Ddx18 protein); C.3′UTR negative-strand by RNA pull down of HA-Ddx18 (M.180 ku protein marker; lane 1. input; Lane 2.Blank; lane 3.Bio-ds-3′UTR RNA; Lane 4.Bio-ss(-) 3′UTR); D. Ddx18 protein by RIP pull-down of the 3′UTR negative-strand; E. Purified Ddx18 protein with His tag (M.180 ku protein marker; Lane 1.Blank; Lane 2.purified Ddx18 protein with His tag); F. Western blot verification of normal expression of the purified His-Ddx18 protein (M.180 ku protein marker; Lanes 3 and 4.Ddx18 purified protein with His tag); G. Verification of direct interaction of the 3′UTR negative-strand with His-Ddx18 protein by EMSA assay"

Fig. 8

3′UTR predicted structure and truncation diagram and validation of Ddx18 and 3′UTR interaction region A. Schematic diagram of single negative-strand 3′UTR RNA secondary structure; B. Schematic diagram of single negative-strand 3′UTR truncation; C. RNA pull down to probe the region of Ddx18 interactions with single negative-strand 3′UTR truncated RNA (M.180 ku protein marker; 1. Bio-ss-3′UTR; 2. Bio-ss-3′UTR-ΔpolyU; 3.Bio-SL1; 4.Bio-SL2)"

Fig. 9

Ddx18 interacts with 2C by immunoprecipitation A. PCR amplification of 2C target fragment (M. DNA relative molecular mass; Lane 1. 2C); B. Double digestion of p3×FLAG-2C plasmid (M. DNA marker; Lane 2. p3×FLAG-2C); C. Western blot verification of 2C protein expression (M.180 ku protein marker; Lane 1.2C protein with flag tag); D.Ddx18 immunoprecipitation with 2C"