SUMO-2/3 enrichment in stabilizing gene transcriptional during KSHV reactivation. (A) TREx-F3H3-K-Rta-shSUMO-2/3 BCBL-1 cells were treated with Dox for 24 and 48 hours. TCLs were analyzed by immunoblotting using anti-SUMO-2/3 antibody. (B) Twelve IRF-1, IRF-2 and IRF-7 targeted genes showing SUMO-2/3 enrichment at the promoter region during KSHV reactivation were chosen. Two genes showing no SUMO-2/3 enrichment at the promoter region were chosen as control. RNA samples derived from TREx-F3H3-K-Rta BCBL-1 and TREx-F3H3-K-Rta-shSUMO-2/3 BCBL-1 cells before and after 24 hours of Dox induction were subjected to reverse transcription (RT) reaction. Following the RT reaction, the IRF target genes were amplified by qPCR using gene-specific primer sets. All reactions were run in triplicate and normalized against GAPDH.