Figure 1 | Scientific Reports

Figure 1

From: DUSP6 is a memory retention feedback regulator of ERK signaling for cellular resilience of human pluripotent stem cells in response to dissociation

Figure 1

Cell passaging with EDTA increases DUSP6 expression. (A) Illustration depicting the experimental setup. hPSCs were passaged using the EZPassage tool for mechanical passaging (Mech) and 0.5 mM EDTA in DPBS for chemical passaging (EDTA). After three passages, hPSCs were harvested and RNA-seq analysis was performed. Clumps obtained after dissociation by each method are shown (scale bar, 1 mm). Samples were used in duplicates. (B) Differentially expressed genes (DEGs) in hFSiPS1 and SNUhES31 cells passaged using the mechanical and chemical methods. DEGs were selected on the basis of fold change (> 1.8) and p-values (< 0.05) (C) Heatmap of DEGs. Genes associated with MAPK signaling are marked in red. (D) Gene Ontology analysis of DEGs revealed that genes regulating the MAPK signaling pathway are significantly enriched. (E) Evaluation of DUSP6 mRNA expression in mechanically and chemically passaged cells (n = 6) by real-time PCR. (F) Levels of DUSP6 protein were analyzed by western blotting.

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