Figure 5

Oxidative stress is involved in hematopoietic deficiencies caused by GRK6 ablation. (a) Network analysis of over-represented pathways in RNA-seq. Each node represents a pathway, its size correlates with enrichment significance and edges represent gene overlaps between nodes. Network clusters are indicated by grey shadows. (b) Representative flow cytometric analysis and statistics of ROS level assessed by DCF-DA fluorescence (n=5, 3). *P<0.05 by two-tailed, unpaired Student’s t-test. (c) WT and GRK6^−/− mice was randomized to i.p. injection of saline or LA (50 mg/kg) for two weeks (WT+Saline, WT+LA, GRK6^−/−+Saline, GRK6^−/−+LA, n=5, 4, 3, 3). HSC, two-way ANOVA, with Bonferroni post hoc, P=0.01, WT versus GRK6^−/−, P=0.305, LA versus Saline. CLP, two-way ANOVA with Bonferroni post hoc, P=0.015, WT versus GRK6^−/−; P=0.874, LA versus control; **P<0.01, ***P<0.001, within genotype; ^#P<0.05, ^###P<0.001, within treatment. (d) Methocellulose-based CFU-C and CFU-Pre-B assay at different LA dosage. CFU-C, two-way RM ANOVA with Bonferroni post hoc; P=0.107, WT versus GRK6^−/−; P<0.001, LA dosage. CFU-Pre-B, two-way RM ANOVA with Bonferroni post hoc, P=0.091, WT versus GRK6^−/−; P<0.001, LA dosage. *P<0.05, **P<0.01, ***P<0.001, Bonferroni post hoc within genotype; ^#P<0.05, ^###P<0.001, Bonferroni post hoc within treatment (versus LA dose 0). Data are expressed as mean±S.E.M.