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. 2022 Apr 19;23(9):4479. doi: 10.3390/ijms23094479

Figure 6.

Figure 6

IRE1 pathway was activated in PRV single- and coinfection groups. PK-15 cells were single- or co-infected with PCV2 and/or PRV for 4, 8, 12, 24, 36, and 48 h. *, p-value < 0.05; **, p-value < 0.01; ****, p-value < 0.0001. (A) Western blotting of p-IRE1, IRE1, EDEM1, and viral proteins PCV2 Cap and PRV gD. β-actin was used as a control. (B) The ratio of p-IRE1α and IRE1α. Quantification of the bands corresponding to the p-IRE1α and IRE1α by densitometry was normalized to β-actin. (C) xbp1 mRNA levels quantified by real-time RT-PCR. (D) Splicing of xbp1 mRNA. Spliced xbp1, sxbp1; unspliced xbp1, uxbp1. (E) EDEM1 mRNA levels were quantified by real-time RT-PCR. (F) The relative amount of EDEM1 to β-actin was quantified by densitometry analysis using the ImageJ software. Phospho-IRE1 (Ser724) (Affinity, Jiangsu, China, 1:1000), IRE1 (Affinity, Jiangsu, China, 1:1000), EDEM1 (Affinity, Jiangsu, China, 1:2000), PRV-gD (Lvdu, Shandong, China, 1:500), PCV2-cap [31] and β-actin (Proteintech, Wuhan, Hubei, China, 1:10,000) were used as primary antibody, respectively. Unprocessed original images can be found in Supplementary Figures S3 and S4.