Fig. S6.

Tcf7 shRNA specifically decreases expression of Tcf1 protein and Tcf7 mRNA. (A) The murine lymphoma line EL4, which constitutively expresses high levels of Tcf1 protein, was transduced with control lentivirus or lentivirus expressing Tcf7 shRNA. Transduced GFP⁺ cells were isolated by FACS sorting and whole cell lysates were prepared, separated by SDS/PAGE, and transferred to PVDF membranes. Tcf1 expression was detected by Western blot, and then the membrane was stripped and reprobed to measure Actin expression. One of three experimental replicates with representative results is shown. (B) DNMT3a KO OT1 T cells were activated in vivo by VacOva infection, isolated after 48 h, and transduced with control or Tcf7 shRNA expressing lentivirus in vitro. GFP⁺ transduced cells were isolated by FACS sorting after 48 h and RNA was extracted and converted to cDNA. Expression of Hprt and Tcf7 transcripts was measured by Taqman qPCR using 18S RNA expression to normalize input. Target gene expression is normalized to that of control transduced cells. Plots depict mean ± SD of triplicate technical replicates.