ETS2 and ETS1, which are downstream targets of miR-221 and miR-222, respectively, induce EC migration. (A) ETS2 is repressed by miR-221. The expression level of ETS2 was detected by qRT-PCR (top panel) or immunoblotting (bottom panel) in HMEC1 cells with stably expressed miR-221. (B) ETS1 is repressed by miR-222. The mRNA and protein levels of ETS1 was detected in HMEC1 cells with stably expressed miR-222. (C-D) ETS2 and ETS1 are up-regulated in KSHV-infected LECs and BECs. The expression level of ETS2 (C) or ETS1 (D) was detected in KSHV-infected LECs or BECs. (E) ETS2 and ETS1 are up-regulated in KSHV-infected HMEC1 cells. The protein levels of ETS2 and ETS1 were detected by immunoblotting in HMEC1 cells or HK cells. Relative band intensities are shown. (F-G) Both ETS2 and ETS1 are able to induce EC migration. HMEC1 cells stably transduced with an ETS2-expressing plasmid (F) or an ETS1-expressing plasmid (G) were used to detect their protein expression level by immunoblotting (left panel) and were also subjected to the Transwell cell-migration assay (right panel; n = 3). (H) Structure of the ETS2 transcript and predicted seed region duplex formed between ETS2 and miR-221 (left panel). Reporter activity of the ETS2 3′UTR reporter construct after cotransfection with the miR-221–expressing or empty vector into 293T cells (right panel; n = 3). (I) Structure of the ETS1 transcript and 2 putative miR-222–binding sites (top panel). Reporter activity of the ETS1 3′UTR reporter constructs (UTR1 or UTR2) after cotransfection with the miR-222–expressing or empty vector into 293T cells (bottom panel; n = 3).