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. 2020 Aug 10;16(8):e1008730. doi: 10.1371/journal.ppat.1008730

Fig 6. vIRF1 upregulates VEGFA expression by activating the SPAG9/JNK1/2 pathway.

Fig 6

(A). Western-blotting analysis of SPAG9, phosphorylated JNK1/2, total JNK1/2 and VEGFA expression in HUVECs transduced with lentiviral-vIRF1or its control lentiviral-pHAGE. (B). Western-blotting analysis of phosphorylated JNK1/2, total JNK1/2 and VEGFA expression in vIRF1-expressing HUVECs transduced with lentivirus-mediated No.1 (shSPAG9-1) and No. 2 (shSPAG9-2) shRNAs targeting SPAG9. (C). Western-blotting analysis of phosphorylated JNK1/2, JNK1/2 and VEGFA in vIRF1-expressing HUVECs treated with the JNK inhibitor, SP600125 (50 μM) for 48 h. (D). Western-blotting analysis of phosphorylated JNK1/2, JNK1/2 and VEGFA in KSHV-infected HUVECs treated with the JNK inhibitor, SP600125 (50 μM) for 48 h. (E). Western-blotting analysis of SPAG9, phosphorylated JNK1/2, JNK1/2 and VEGFA expression in iSLK-RGB cells treated with doxycycline (Doxy) (1 μg/ml) for 48 h. (F). Western-blotting analysis of SPAG9, phosphorylated JNK1/2, JNK1/2 and VEGFA expression in Doxy-induced iSLK-RGB cells transduced with lentivirus-mediated No.1 (shSPAG9-1) and No. 2 (shSPAG9-2) shRNAs targeting SPAG9. (G). Luciferase reporter assay of the activity of VEGFA promoter in HUVECs transduced with lentiviral-vIRF1 or its control lentiviral-pHAGE. (H). Luciferase reporter assay of the activity of VEGFA promoter in vIRF1-expressing HUVECs treated with the JNK inhibitor, SP600125 (50 μM) for 48 h. (I). Western-blotting analysis of phosphorylated JNK1/2, JNK1/2 and VEGFA expression in vIRF1-expressing HUVECs transduced with lentivirus-mediated a mixture of shRNAs targeting Lef1 (shLef1). (J). Luciferase reporter assay of the activity of VEGFA promoter in vIRF1-expressing HUVECs transduced with lentivirus-mediated a mixture of shRNAs targeting Lef1 (shLef1). The quantified results represent mean ± SD. * P < 0.05, ** P < 0.01, and *** P < 0.001, Student's t-test.