Extended Data Fig. 8. (Related to Sub-heading, Persistence is linked to seronegativity).Presence of CAN-3110 DNA in un-injected temporal lobe lesion of subject in Fig. 3b (8 months from injection in occipital lesion).

(a) Schematic of CAN-3110 viral genomic DNA showing the location of primers for Nestin-Hsp68 relative to other transcriptional cassette elements. (b) Gel electrophoresis of the PCR products from genomic DNA extracted from FFPE of postmortem brain from subject 014. Primers for Nestin-Hsp68 amplified a PCR product of ~112 bps within the promoter/enhancer transcriptional cassette. WT HSV represents a negative control with viral genomic DNA from HSV1 strain 17+. CAN-3110 represents a positive control with CAN-3110 viral genomic DNA amplifying a strong band of ~112 bp in the Nestin-Hsp68 promoter PCR reaction. BA-19-036 is the temporal lobe tumour that was not injected with CAN-3110 but was IHC-positive for HSV antigen (see Fig. 3b). The uncropped version of this gel is shown in Supplementary Fig. 1. Each PCR reaction was run in triplicate. (c) Sequencing reactions of CAN-3110 control and BA-19-036 PCR products show sequence homology with the original CAN-3110 sequence map.