American Society of Hematology

Figure 2

$Figure 2. GSK3β/beta-catenin phosphorylation and levels in UCB-derived CD34+ cells cultured in 2% to 3% O2 on PLL- versus FN-coated plates. (A) Integrin-preactivated CD34+ progenitors from different cord blood donors were pooled and plated at 1 million cells per 3 mL medium containing SCF, FLT3L, TPO, on PLL- versus FN-coated plates for 48 hours at 2% to 3% O2. Viable cell numbers were normalized by Trypan blue count and then subfractionated for protein analysis. Immunoblots were probed with antibodies against beta-catenin, phospho-Ser9-GSK3β, phospho-Tyr216-GSK3β. Blots were stripped and reprobed with GSK3β antibody. (B) Upper part of the blots (higher than 70 kDa) was immunoblotted with antibodies against HIF-1 alpha. All blots were reprobed with antibodies against laminin and tubulin, which served as nuclear and cytoplasmic control, respectively.$

GSK3β/beta-catenin phosphorylation and levels in UCB-derived CD34⁺ cells cultured in 2% to 3% O₂ on PLL- versus FN-coated plates. (A) Integrin-preactivated CD34⁺ progenitors from different cord blood donors were pooled and plated at 1 million cells per 3 mL medium containing SCF, FLT3L, TPO, on PLL- versus FN-coated plates for 48 hours at 2% to 3% O₂. Viable cell numbers were normalized by Trypan blue count and then subfractionated for protein analysis. Immunoblots were probed with antibodies against beta-catenin, phospho-Ser9-GSK3β, phospho-Tyr216-GSK3β. Blots were stripped and reprobed with GSK3β antibody. (B) Upper part of the blots (higher than 70 kDa) was immunoblotted with antibodies against HIF-1 alpha. All blots were reprobed with antibodies against laminin and tubulin, which served as nuclear and cytoplasmic control, respectively.

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