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. 2017 Feb 17;13(2):e1006216. doi: 10.1371/journal.ppat.1006216

Fig 4. JMJD2A SUMOylation is crucial for SUMO-2/3 enrichment on the KSHV genome during viral reactivation.

Fig 4

(A) TCLs from TREx-MH-K-Rta-shJMJD2A-Flag-JMJD2A-WT and -K471R BCBL-1 cells before and after 12 hrs Dox (0.2 μg/ml) treatment were immunoblotted with antibodies as indicated. Ratio is the relative signal of K-Rta or K-bZIP to α-Tubulin observed for Dox treatment at 24 hrs using TREx-MH-K-Rta-shJMJD2A-Flag-JMJD2A-WT BCBL-1 cells as 1.0. (B) ChIP-seq assay was performed with chromatin from cells treated as described in (A) using rabbit IgG and anti-SUMO-2/3 antibodies. Sequenced reads mapped to KSHV genome were normalized with total reads from the human genome. The histogram shows reads per million mapped reads (RPM) mapped across the KHSV genome (JMJD2A-WT, blue; -K471R, red). p = 2.27e-103 by Student’s-t test. The blue rectangles are high SUMO-2/3 regions as in Fig 1A. (C) ChIP-seq data was verified by real-time qPCR using primer pairs specific for promoter regions of KSHV K6, PAN, K-bZIP, Orf52, Orf23 and Orf25. (Data represent mean±SEM. n = 3. **p<0.01. ***p<0.005).