(A) Primary HDB and GBM tissue was subject to enzymatic dissociation followed by plating in serum-free adherent culture. Representative IF of key lineage markers in primary HDB and GBM dissociated adherent cultures is shown (see also Figure S2C).
(B) Representative IF (left) and RT-qPCR (right) of 6 HDB and 20 GBM dissociated adherent cultures. Highly refractory and moderately refractory GBM cultures were defined with reference to the median ZIKV copy number for all GBMs analysed (ZIKV MOI:1 48 h and ZIKV MOI:3 72 h respectively). Each RT-qPCR data point represents the average copy number across three replicate wells/slices per sample. (Mann-Whitney test *** = p <0.001, **** = p<0.0001)
(C) Representative IF (left) and % cells SOX2+ in HDB (n=4 specimens), HR GBM (n=4 patients) and MR GBM (n=4 patients) adherent cultures (left) and. Cell counts for IF and smFISH determined by manual counting of at least 200 cells per condition. Mean values across all biological replicates for each condition are indicated by horizontal lines. (ANOVA p< 0.0001)