Fig. 1

Human CML and B-ALL patients derived stem cells and progenitors (HSC/P) express aPKCι predominantly. a Representative example of western blot analysis of BCR-ABL, phospho-aPKC, aPKCι, aPKCζ, and actin in HSC/P (CD45⁺/CD34⁺) derived from healthy donors and CML patients mobilized peripheral blood (PB). 5 μg of whole cell lysate was loaded into each lane. b Western blot band intensity ratio between phospho-aPKCι and total aPKC expression. Atypical protein kinase Cι is predominantly expressed, and CML CD34⁺ cells show an increased level of activated aPKCι (phospho-aPKC) in comparison to healthy donor-derived CD34⁺ cells. c, d Histogram plot (c) and quantitation of phospho-aPKCι (d) as evidenced by phospho-flow analyses in B progenitors (CD45⁺/CD34⁺/CD19⁺) derived from healthy donor BM (n = 2) and B-ALL patients (n = 3). B-ALL patient’s derived B-cell progenitors show enhanced p-aPKCι level in comparison to healthy donor-derived counterparts. e Representative example of western blot analysis of BCR-ABL, phospho-aPKCλ, aPKCλ, aPKCζ, and actin in mock (empty vector) and BCR-ABL-transduced murine HSC/P. f Western blot band intensity ratio between phospho-aPKCλ and total aPKC expression. g Representative western blots of BCR-ABL, p-aPKCλ, aPKCλ, and actin in the input and α-aPKCλ antibody immunoprecipitated (IP) fraction. Western blots with different exposure time for aPKCλ and phospho-aPKCλ were presented for quantification accuracy; Low (15 sec), high (1 min). Cont: control. Data are presented as mean ± SD of a minimum of 2 independent experiments. *p < 0.05; **p < 0.01, t-test