FIG 6.

KD of METTL3 decreases m⁶A and reduces KSHV lytic gene expression. (A) Levels of METTL3 mRNA in BCBL1 cells expressing METTL3-specific shRNA from Santa Cruz Biotechnologies (shRNA-SC) or Origene Technologies, Inc. (shRNA-OT), or control shRNA. The cells were treated with PBS or TPA for 24 h. (B and C) Levels of ORF50 (RTA) (B) and ORF57 (C) mRNA in the cells described in the legend to panel A. (D) Levels of m⁶A-mRNA of ORF50 (RTA) in the cells described in the legend to panel A. (E) Western blot detection of METTL3 and KSHV lytic proteins encoded by ORF50 (RTA) and ORF57 in the cells described in the legend to panel A. The level of β-tubulin was used as a loading control. (F) Relative levels of KSHV DNA in the supernatants of the cells described in the legend to panel A at 96 h after TPA stimulation, which were determined as described in the legend to Fig. 5F.