Fig. 2. Adipocyte-secreted factors promote cellular aggregation and reduce the proliferation of human B-ALL cells.
a REH and SEM B-ALL cell lines were cultured in unconditioned medium (RPMI), SCM, or ACM. Brightfield microscopy images were taken after 3 days and cluster sizes were determined by ImageJ analysis. b Human B-ALL cell lines (Nalm6, REH, SEM, RCH-AcV, and 697) were plated in RPMI, SCM, or ACM at a starting density of 5 × 105 cells/well (indicated by the dotted line). The number of viable cells was determined on day 3 of culture using trypan blue exclusion assays. c The percentage of apoptotic or dead cells in b was determined using Annexin V/PI assays and analyzed using flow cytometry. d Human B-ALL cell lines were cultured in RPMI, SCM, or ACM for 24 h. Lysates were harvested and β-Galactosidase activity was determined using the Senescence β-Galactosidase Activity Assay Kit (Cell Signaling Technology) per the manufacture’s protocol. e Human B-ALL cell lines (REH and 697) were treated with RPMI, SCM, or ACM for 24 h and the protein levels of AKT and ERK (phospho- and total) were determined via western blot analysis. Means + s.d. are shown for a, b, c and d. *P < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001, one-way ANOVA with Tukey’s post-test, n = 3 independent experiments for a, b, c, and d. A representative of n = 3 experiments is shown in e. Western blot source data are provided in the Source Data file.