Fig. 1

PFV infection upregulates SAP30. (A) HT1080 cells were infected with PFV for 6 h, 12 h, and 24 h. Total RNA was extracted for transcriptomics sequencing. GO enrichment analysis from the PPI network of differentially expressed genes generated via the STRING database. The top 10 enriched GO biological processes are shown, with the second-ranked term, “negative regulation of RNA polymerase II-mediated transcription,” highlighted in red. (B) The result of transcriptomic sequencing. (C) HT1080 cells were infected with PFV for 6 h, 12 h, and 24 h. Total RNA was extracted and reverse transcribed into cDNA. Real-time PCR was performed with gene-specific primers to detect the mRNA level of SAP30. (D) HT1080 cells were transfected with pGL-3162 (a recombinant plasmid containing the SAP30 promoter) or pGL3-basic (a luciferase reporter vector lacking promoter sequences, function as a negative control). Twenty-four hours post-transfection, cells were infected with PFV (MOI = 0.25 or 0.5). Luciferase activities were measured and normalized to β-gal catalytic activities. (E) HEK293T cells were transfected with pGL-3162 or empty vector, along with Flag-Env, Flag-Gag, Myc-Bet, 3.1-Tas and pCMV-β-gal. Forty-eight hours post-transfection, luciferase activities were measured and normalized to β-gal catalytic activities. (F) HEK293T cells were transfected with Flag-Tas or empty vector, and SAP30 expression was detected by Western blotting 48 h post-transfection