Skip to main content
. 2026 Apr 21;100(5):e00119-26. doi: 10.1128/jvi.00119-26

Fig 8.

SHFL domain schematic with mutants and frameshifting efficiency data in BHK-21 and HEK-293T cells. Western blots show protein expression levels identifying amino acids 164-199 as crucial for PRRSV -1 PRF inhibition.

Identification of the key region essential for SHFL’s inhibitory effect on PRRSV −1 PRF. (A) Schematic diagram of SHFL domains and mutants. Deleted regions are represented by dashed lines; Mutation regions are highlighted in red; Key point mutations are marked by asterisks (*); Δ indicates the lack of the denoted region. (B–G) The 164th–199th amino acid region of SHFL is a key region involved in suppressing PRRSV −1 PRF. The −1 PRF reporter plasmid was cotransfected with different SHFL mutants in BHK-21 (B and C) and HEK-293T (D and E) cells. (F and G) The −1 PRF reporter plasmid was cotransfected with SHFL-3R/A mutant in HEK-293T cells. In all experiments, cell lysates were harvested at 24 hpt and analyzed by dual-luciferase assay and Western blot using an anti-FLAG antibody to detect SHFL mutant expression. β-Tubulin was detected as a loading control. (B, D, and F) Data are presented as means ± SD from three replicates. Statistical significance was analyzed using the two-tailed unpaired Student’s t-test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.