FIG 3.

C19ORF66 mRNA is protected from herpesviral endonucleases by its 3ʹ UTR and accumulates in the cytoplasm of iSLK.219 cells. (A) 293T cells were transfected with a GFP reporter (GFP) or a GFP reporter containing C19ORF66 3ʹ UTR sequence along with a control empty vector (mock) or a plasmid expressing SOX, muSOX, BGLF5, or vhs. After 24 h, total RNA was harvested and subjected to RT-qPCR to measure GFP mRNA levels. (B) KSHV-positive iSLK.219 cells were reactivated for the indicated times to induce KSHV lytic cycle (lyt) or not (KSHV latent phase maintained [lat]). Cells were harvested, lysed, resolved on SDS-PAGE, and Western blotted with the indicated antibodies. (C) Unreactivated (lat) or reactivated (lyt) KSHV-positive iSLK.219 cells were fractionated into nuclear and cytoplasmic fractions and Western blotted with the indicated antibodies.