a. Immunoblot analysis revealed restored β-catenin protein expression in shβcat-expressing CML CD34+ cells cultured in HS-5 DC (left, n=3). qRT-PCR confirmed β-catenin knockdown at the mRNA level (right, n=3). Immunoblots were normalized to α-tubulin and qRT-PCR was normalized to GUS.
b. Consistent with restored β-catenin protein levels, HS-5 DC also restored colony forming ability of shβcat-expressing CML CD34+ cells under scenarios of DC in the presence of imatinib (n=4). c. Addition of an N-cadherin blocking antibody to co-cultures restored the effects of imatinib and shβcat on colony formation of CML CD34+ cells grown in HS-5 DC (n=3). d,e. Addition of antibodies blocking N-cadherin (n=9), H-cadherin (n=8), or both (n=6), to HS-5 co-cultures restored the effects of imatinib on CML CD34+ cells cultured under HS-5 DC in the presence of imatinib (d), but did not have an effect on β-catenin protein levels as demonstrated by immunoblot analyses (e). Bars represent SEM. *p<0.05.