Fig. 5

HCMV-miR-US33-5p downregulates SLC3A2 by targeting EPAS1. A RNA sequencing data showing EPAS1 differential expression between the HCMV-miR-US33-5p and control groups. B, C HCMV-miR-US33-5p was transfected into HA-VSMCs. qRT-PCR (B) and Western blot (C) were used to analyze the EPAS1 expression level in HA-VSMCs. Western blot was performed three times; representative images are presented. Numbers above lanes represent the relative protein level, normalized relative to control samples. D Potential EPAS1 binding sites in SLC3A2 promoter region. E Luciferase reporter assay of the effect of EPAS1 on the SLC3A2 promoter. F ChIP assay showed that EPAS1 bound to SLC3A2 promoter. G ChIP-qPCR assay of the enrichment of EPAS1 to SLC3A2 promoter. H Potential HCMV-miR-US33-5p binding site in EPAS1 promoter. I Luciferase reporter assay of the effect of HCMV-miR-US33-5p on the EPAS1 promoter. J RIP analysis of the enrichment of Ago2 on 3′-UTR of EPAS1 and HCMV-miR-US33-5p. **P < 0.01, ***P < 0.001 (n = 3, ANOVA analysis for A and D; Student’s t-test for F)