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

Fig 3.

Western blots, fluorescence images, and virus titer graphs show SHFL knockout enhances PRRSV infection. Data reveal higher viral protein expression and increased viral replication in SHFL-KO cells across three viral strains at multiple time points.

SHFL knockout enhances PRRSV infection irrespective of viral species. (A) Polyclonal MARC-145 cells with SHFL knockout by CRISPR/Cas9 gene editing. Endogenous SHFL expression in WT and SHFL-KO MARC-145 cells treated with IFN-α2b (0, 10, and 100 IU/mL) for 12 h was evaluated by Western blot analysis. (B–D) SHFL knockout enhanced the infection of rTA-EGFP. WT and SHFL-KO MARC-145 cells were infected with rTA-EGFP (MOI = 0.1). The infection of TA-EGFP in WT and SHFL-KO MARC-145 cells at the indicated time points was assessed by green fluorescence signal (B; scale bar, 100 μm), Western blot analysis of viral protein expression (C), and virus titers in culture supernatants (D). (E) One-step growth curves of rTA-EGFP in WT and SHFL-KO MARC-145 cells. Cells were infected with rTA-EGFP (MOI = 3). At the indicated times, supernatants were harvested and subjected to viral titration in MARC-145 cells. (F and G) SHFL knockout enhanced the infection of the NL1207 strain and the SHE strain. WT and SHFL-KO MARC-145 cells were inoculated with the NL1207 strain (MOI = 0.1) or the SHE strain (MOI = 0.1). At the indicated times, culture supernatants were harvested for virus titration. (D–G) Data presented are means ± SD from three replicates. Statistical significance was analyzed using multiple unpaired t-tests. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant.