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. 2023 Mar 6;97(3):e00011-23. doi: 10.1128/jvi.00011-23

FIG 5.

FIG 5

ORF8 induces stress-like responses in human cells. (A) ORF8 induces HSPA5 upregulation. ORF8-Flag (ORF8 +) or Flag (ORF8 -) was overexpressed in human cells, as indicated for 60 h, and cell lysates were analyzed by WB. (B) ORF8 induces sustained XBP-1 splicing. RNAs obtained from ORF8-overexpressing or Tg-treated 293T cells at the indicated time posttransfection/treatment were analyzed for XBP-1 splicing detection. (C) Transcriptome abnormalities induced by ORF8. Log2-fold changes (fold change) of gene expression (ORF8 positive versus ORF8 negative) and -log10p value (P value) are shown, including genes with various expression levels in the yellow sector in upper left: high (100 ≤ average FPKM), moderate (10 ≤ average FPKM < 100), low (1 ≤ average FPKM < 1) and extremely (ex) low (average FPKM < 1). (D-F) ORF8 induced upregulation of HSPA5, PDIA4 and CHOP effectors of the stress-like response. RNAs from ORF8-overexpressing 293T cells at the indicated times posttransfection were analyzed by qRT–PCR, and the target genes are shown in the upper left. Significance: *(P ≤ 0.05), **(P ≤ 0.01), and ***(P ≤ 0.001).