Fig. 4. Adipocyte-secreted factors induce significant gene expression changes in human B-ALL cells in the absence and presence of methotrexate.

Human B-ALL cell lines (REH and RCH-AcV) were cultured with ACM for 48 h or preconditioned with ACM for 24 h and cultured with MTX for an additional 24 h (48 total hours of culture). After 48 h of culture, B-ALL cells were harvested, and RNA isolated for RNA-sequencing analysis. a–d, RNA-sequencing results were analyzed using principal component analysis (PCA). e–f, Unsupervised hierarchical clustering analysis and heatmaps of the RNA-sequencing results are shown with genes upregulated in ACM indicated in black (along with red arrow) and genes downregulated in ACM indicated in gray (along with the green arrow). Treatment conditions are color-coded at the top of the heatmap.