Fig. 2: Small leukemic cells exhibit reduced F-actin and stiffness, leading to increased stemness.
a Schematic of the AFM setting for cell cortical stiffness measurement. b Representative immunofluorescence images of F-actin (red) and nuclei (blue) in small and large AML BM Lin- cells, with quantification of F-actin intensity (n = 113 cells for small, and n = 191 cells for large; from four AML samples). c AFM-based stiffness measurement (n = 23 cells per group, from four AML samples). d Correlation between cell size and stiffness (n = 46 cells, from four AML samples). Imaging flow cytometry of F-actin (e) and the cell area (f) for Actinlow and Actinhigh cells from six AML samples. g–j F-actin staining (g, h, n = 5 AML samples) and stiffness quantification (i, n = 19 individual cells per group, from five AML samples) in small and large primary BM Lin- cells treated with 1 μM Jas or 1 μM Lat A for 6 h. Colony formation assay (j, n = 3 AML samples) of treated and untreated AML BM Lin⁻ cells (k, Overview of RNA-seq, in vivo xenografts, and drug sensitivity assays using size-sorted AML BM Lin⁻ cells treated with Jas or Lat A (n = 5 AML samples). l DEGs between small and large cells. m GSEA of DEGs between small and large cell. n DEGs between untreated and Jas-treated small cells. o GSEA of DEGs between untreated and Jas-treated small cells. p DEGs between untreated and Lat A-treated large cells. q GSEA of DEGs between untreated and Lat A-treated large cells. r Flow cytometry analysis of BM engraftment following transplantation of treated AML BM Lin⁻ cells (n = 4 mice per group, from three AML samples). s Cell viability assay following Ara-C treatment (48 h) in size-sorted AML BM Lin⁻ cells pretreated with Jas or Lat A (n = 5 AML samples). For (b, c, s), data were analyzed using two-tailed unpaired Student’s t test. For (d), data were analyzed using Pearson correlation. For (f, l–q), data were analyzed using two-tailed paired t test. For (h–j, r), data were analyzed using one-way ANOVA with Tukey correction (F = 38.13, 41.74, 24.37 and 56.32, respectively). Data are mean ± SEM and representative of at least three independent experiments. Source data are provided as a Source Data file.
