GSK3β is highly upregulated, active, and more nuclear in AML cells. (A) GSK3β levels were assessed as indicated in normal bone marrow (NM), immature granulocyte colony-stimulating factor mobilized peripheral blood leukocytes (MB), and AML cells by western blot. FAB subtypes are as follows: HL60-M2 OCI-M4 U937-M4 subtype THP1-M5 subtype Pt1-M4 Pt2-M2 Pt3-M4EO. (B) GSK3β levels in different populations of normal cells (n = 2 for each subpopulation) and AML cells (n = 2 for 3 cell lines and 1 patient sample) were analyzed using flow cytometry analysis. P < .001 for all comparisons except for OCI/CD3, Pt/CD3, OCI/CD19, Pt/CD19, OCI/CD14, and Pt/CD14 where P < .01. (C) Imaging cytometry was used to quantify GSK3β expression in primary AML (n = 86) and normal bone marrow CD34⁺ cells (n = 12) expressed in box-and-whisker plot displaying the median and quartile distributions of GSK3β mean fluorescence intensity. (D) Imaging cytometry was used to quantify GSK3β nuclear localization in primary AML (n = 86) and normal bone marrow CD34⁺ cells (n = 12) expressed in box-and-whisker plot displaying the median and quartile distributions. (E) Representative images of cells from the cell types indicated from imaging cytometry showing GSK3β expression and the nucleus (4′,6-diamidino-2-phenylindole [DAPI] staining). The GSK3β expression level and nuclear localization of each sample is shown. **P < .01; ***P < .001.