Figure 3.

G596R and L597V, but not V600E, mutant BRAF are overexpressed in melanoma recurrences. cDNA was analyzed by qrtPCR for expression of BRAF from primary tumors treated with PBS, or early (0.2–0.3 cm diameter) explant recurrent tumors derived from primary B16tk (a–c) or B16ova (d), tumors in C57BL/6 mice induced to regress, enter a state of MRD and then recur, following (a) GCV chemo,¹ (b) paclitaxel/reovirus,²⁶ (c) VSV-cDNA ASMEL immune,⁷ or (d) (B16ova) OT-I adoptive T cell^8,9 frontline therapies (as described in Materials and Methods). The threshold cycle (Ct) at which amplification of the target BRAF sequence was detected was used to compare the relative levels of mRNA between samples. CT BRAF-GAPDH: relative quantities of BRAF mRNA were normalized with Ct of GAPDH amplification. (e) cDNA was analyzed by qrtPCR for expression of BRAF from primary TC2 tumors treated with PBS, or early (0.2–0.3 cm diameter) explant recurrent tumors derived from primary TC2 tumors in C57BL/6 mice induced to regress, enter a state of MRD and then recur, following VSV-cDNA ASEL immune frontline therapy (as described in Materials and Methods).²⁴