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. 2018 Feb 15;103(4):666–678. doi: 10.3324/haematol.2017.182048

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Figure 1. — Identification of differentially regulated genes in adherent co-culture cells. (A) A fraction of co-cultured Jeko-1 cells adhere to an MS-5 cell mono-layer and remain stably bound in the presence of excess suspension cells. The number of unlabeled (light gray) or CFDA-SE labeled (dark gray) Jeko-1 cells adhered to MS-5 stromal cells after 24 hours (h) was not significantly reduced after a further 24 h of incubation (48 h) with excess CFDA-SE (P=0.841) or unlabeled (P=0.391) Jeko-1 cells, respectively. Mean±Standard Deviation are shown for cultures (n=3). (B) Experimental design showing the three Jeko-1 and the two MS-5 stromal cell fractions for which RNAseq data were acquired. (C) Heatmap representation of relative transcript levels for the 3697 genes with significantly changed mRNA levels (FDR q-value ≤ 0.05) in at least one Jeko-1 cell fraction after hierarchical clustering. (D) Venn diagram illustrating number of differentially expressed genes in pair-wise comparisons between the three Jeko-1 cell fractions (FDR q-value ≤ 0.05, total number of changed genes: 3697). The three comparisons in the Venn diagram are: adherent Jeko-1 cells in co-culture (ADH) compared with suspension Jeko-1 cells in co-culture (SUSP), n=1050 (ADH/SUSP), ADH compared to mono-cultured Jeko-1 cells (SEP), n=3453 (ADH/SEP) SUSP compared to SEP n=1471 (SUSP/SEP). (E) Ranked fold changes of 1050 genes with changed transcript levels in ADH relative to SUSP Jeko-1 cells in co-culture.