Extended Data Fig. 6: Corin induces EBV lytic reactivation in EBV⁺ transformed Burkitt and lymphoblastoid B-cells and in Burkitt xenograft tumors in vivo.

(A) Corin and ganciclovir co-treatment effects on EBV-negative Burkitt and EBV⁺ LCL cells. Shown are mean ± SD fold changes of live cell numbers, relative to DMSO-treated controls, from n = 3 replicates on day 6 of treatment of EBV-negative MUTU I (left) and of EBV⁺ GM15892 LCLs (right). Cells were treated as described in Fig. 3e with DMSO or corin (0.5 μM). (B) Corin and ganciclovir co-treatment effects on KEM III LCLs. KEM III LCLs were treated with a single dose on day 1 of or with multiple drug doses as described in Fig. 3e. Shown are mean ± SD fold changes of live cell numbers, relative to DMSO-treated controls, from n = 3 replicates on day 6 of treatment. (C) P3HR-1 Burkitt cells were treated with a single dose on day 1 or with multiple doses of DMSO vehicle, LSD1 inhibitor C12 (0.5 μM), the HDAC inhibitor NaB (0.1 mM), C12 + NaB or with the dual LSD1/HDAC inhibitor corin (0.5 μM), using the regimen described in Fig. 3e. Shown are mean ± SD fold changes of live cell numbers, relative to DMSO-treated controls, from n = 3 replicates on day 6 of treatment. (D) Immunoblot analysis of WCL prepared from MUTU I xenografts harvested from mice treated with DMSO vehicle vs corin as described in Fig. 3g. (E) Immunohistochemical analysis of BZLF1 expression in MUTU I xenografts harvested from mice treated with DMSO vs corin, as described in Fig. 3g. Scale bar = 100 μm. Representative images from n = 4 randomly selected fields are presented. (F) Quantification of BZLF1⁺ (left panel) versus BMRF1⁺ (right panel) cell numbers in MUTUI xenograft tumors as in Fig. 3h and Extended Data Fig. 6E. Shown are mean ± SEM numbers of BZLF1⁺ or BMRF1⁺ cells from four randomly selected fields per mouse, quantitated by Image J using the Cell Counter plugin. (G) qPCR analysis of BZLF1, early BMRF1 and late BLLF1 (encodes gp350) mRNA abundances in xenograft tumors from tumors shown in Fig. 3h and Extended Data 6E. Shown are mean of n = 3 or 4 qPCR values. (H) qPCR defined intracellular EBV genome copy number from the xenograft tumors shown in Fig. 3h. Box heights indicate mean of n = 3 or 4 qPCR values. (I) MUTU I xenograft tumor sizes from the vehicle control, corin, GCV or corin and GCV treated mice (n = 6 or 7) shown in Fig. 3j. Data are presented as mean ± standard error of the mean (SEM). Significance was determined by cross-comparison of mean tumor sizes from dual drug treated versus vehicle or single drug treated mice. Blots shown are representative images of n = 3 replicates. A-C, statistical significance was determined using unpaired two-sided t-test; F, Statistical significance was determined by two-tail nested t-test; I, Statistical significance was determined using multiple t-test. *p < 0.05, **p < 0.01, ***p < 0.001, NS, not significant.

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