Bcl 6

LN18 or U87-MG at approximately 70% confluence were transfected with each of PZFN1 and PZFN2 CompoZr^™ Knockout Zinc Finger Nuclease plasmids targeted to BCL6 (Sigma Aldrich, USA), using Lipofectamine^® 3000. After both 24 hours and 72 hours, the medium was collected and the debris and detached cell fraction was pelleted by centrifugation, and the supernatant aspirated. The adherent cells were then harvested and pelleted by centrifugation, or combined with the non-adherent fraction before pelleting by centrifugation. Cell pellets were stored at -80° C. gDNA was extracted from ZFN transfected cell pellets using the Quick-gDNA^™ MiniPrep kit (Zymo Research, USA) as per the manufacturers protocol. gDNA was quantified by fluorometry before PCR amplification of a 381 bp fragment of the BCL6 using Phusion^® High-Fidelity DNA Polymerase, and BCL6 locus forward primer (5’-GAAGAGTTCCTCAACAGCCG-3’), and reverse primer (5’-TTCTGGGATTGTTTCCTTGG-3’), using the following parameters: 1 cycle of 98 °C, 30 s. 30 cycles of 98 °C, 10s; 57 °C, 30 s; 72°C, 10 s. Nuclease activity was detected by CEL1 endonuclease digestion of heteroduplex DNA using the Surveyor^® Mutation Detection Kit (IDT, USA) using the manufacturers protocol.