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. Author manuscript; available in PMC: 2018 Jul 10.
Published in final edited form as: Nature. 2018 Jan 17;553(7689):521–525. doi: 10.1038/nature25438

Extended Data Figure 8. Truncation mutagenesis of the MCP N-lasso or Tri1 N-anchor does not notably affect KSHV DNA replication or gene expression.

Extended Data Figure 8

a, b, Viral genome copy number in cells replicating the wild type (WT), MCP-truncated (a) or Tri1-truncated (b) KSHV. Design of MCP truncations or Tri1 truncations is shown in Figs 3i and 4j, respectively. KSHV lytic replication was induced in cells harbouring the wild-type or the mutated KSHV genome. Total DNA was extracted from cells, and viral genome copy number was determined by real-time PCR. Data are mean ± s.e.m. (n = 3 biologically independent samples). c, d, Viral RNA expression in cells replicating the wild-type, MCP-truncated (c) or Tri1-truncated (d) KSHV. Total RNA was extracted from cells induced for KSHV lytic replication. Viral RNA transcripts were quantified by real-time PCR with reverse transcription and presented as fold changes over RNA level of wild-type virus. Data are mean ± s.e.m. (n = 3 or 4 biologically independent samples). e, f, Expression of viral and cellular proteins in cells replicating the wild-type, MCP-truncated (e) or Tri1-truncated (f) KSHV. Correct sizes of truncated Tri1 were verified by western blotting with an anti-Tri1 antibody as shown in f. Verification of truncated MCP was not carried out owing to the lack of anti-MCP antibody. Experiments were repeated independently twice with similar results.