Figure 4: The absence of miR-193b increases basal and cytokine-stimulated signalling in LT-HSCs. | Nature Communications

Figure 4: The absence of miR-193b increases basal and cytokine-stimulated signalling in LT-HSCs.

From: STAT5-regulated microRNA-193b controls haematopoietic stem and progenitor cell expansion by modulating cytokine receptor signalling

Figure 4

(a) Functional annotation analysis using DAVID (KEGG pathways) of the upregulated and downregulated genes in sorted LSK cells derived from miR-193b−/− and miR-193b+/+ mice determined via RNA sequencing. Only clusters with a score >1 are displayed. (b) Representative histograms and quantification of phosphoflow cytometry analysing basal signalling pathways of HSPCs from miR-193b−/− and miR-193b+/+ mice. N=3 independent experiments. (c) Equal signalling intensities were observed in sorted LT-HSCs after 1 h of starvation (tonic signalling) as assessed by phosphoflow cytometry. N=4 independent experiments. (d) Representative histograms and quantification of phosphoflow cytometry of LT-HSCs stimulated with a myeloid cytokine cocktail for 20 min after starvation. The mean fluorescence intensity (MFI) was normalized to starved and unstimulated cells. N=4 independent experiments. (e) Quantification pSTAT5 and pAKT via phosphoflow cytometry and miR-193b expression via qPCR in LT-HSCs stimulated with a cytokine cocktail at various time points. The miR-193b expression was normalized to snoRNA202. N=3 mice per group (phosphoflow cytometry) and N=3 individual experiments (qPCR). All data are represented as the mean±s.d. *P<0.05; **P<0.01.

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