FIGURE 5.

RNA-binding proteins and production of viral circRNAs. RNA-binding proteins (RBPs) were knocked-down with siRNA pools and circular RNAs and their linear mRNAs counterparts were quantitated. (A,B) KSHV-negative SLK cells were knocked-down for RBPs and RBP-coding transcript levels (n = 4) (A), and hsa_circ_0001400 and RELL1 expression levels (n = 3) (B) were quantitated with RT-qPCR. Expression levels were normalized to non-targeting siRNA (siNT) control. 18 S RNA was used as internal control. (C) iSLK-BAC16 cells were knocked-down for RBPs, reactivated for lytic cycle, and viral circRNAs and their linear counterparts were quantitated with RT-ddPCR. Absolute copy numbers were quantitated and normalized to non-targeting siRNA control. (D) Primary LECs were knocked-down for RBPs, de novo infected for 3 days at MOI of 30, and viral circRNAs and their linear counterparts were quantitated with RT-ddPCR. Absolute copy numbers were quantitated and normalized to siNT control (n = 3). Error bars indicate standard deviation. Statistical significances were calculated by paired ratio t-test. *p < 0.05.