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. 2016 Apr 21;6(5):692–703. doi: 10.1016/j.stemcr.2016.03.006

Figure 3.

Figure 3

cAMP Induction Reduces Oxidative Stress and Induces CXCR4 in hPSC-Derived Hematopoietic Cells

(A) Flow cytometry analysis for detection of reactive oxygen species (ROS) in differentiated hPSC-to-hematopoietic cells at day 14 of differentiation. Representative flow cytometry plots (biexponential x axis) show ROS levels in the hematopoietic surface phenotypes. FMO control, fluorescence-minus-one (staining control).

(B) Quantification of geometric mean fluorescence intensity (gMFI) of CellROX dye as indicated in (A). Data represent mean ± SEM of three independent experiments. Statistical analysis was performed using the t test. Significance compared with the control setting: ∗p < 0.05, ∗∗p < 0.01; n.s., not significant.

(C and D) qRT-PCR expression analysis of the indicated redox-state-regulating genes (C) and p38MAPK-related genes (D) in PSC-derived hematopoietic cells. Relative expression of each gene to housekeeping gene ACTB (β-ACTIN) was calculated, and mean fold change respective to control condition (set at 1) is shown. Data represent mean ± SEM of three independent experiments. Statistical analysis was performed using the t test. Significance compared with the control setting: ∗p < 0.05, ∗∗p < 0.01; n.s., not significant.

(E) Analysis of mature hematopoietic progenitors (CD43+CD34−CD38+) after cAMP induction (day 14). Data represent mean ± SEM of three independent experiments. Statistical analysis was performed using the t test. Significance compared with the control setting: ∗p < 0.05; n.s., not significant.

(F) Expression of CXCR4 across indicated hematopoietic surface phenotypes. Data represent mean ± SEM of three independent experiments. Statistical analysis was performed using the t test. Significance compared with the control setting: ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001; n.s., not significant.

(G) Expression of CXCR4 in HSC-like surface phenotype (ROSlow fraction). Data represent mean ± SEM of three independent experiments. Statistical analysis was performed using the t test. Significance compared with the control setting: ∗∗∗p < 0.001, n.s., not significant.

See also Figure S3.