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. 2015 May 28;4(6):995–1003. doi: 10.1016/j.stemcr.2015.05.003

Figure 1.

Figure 1

Bmi-1 Is Highly Expressed in ESREs and Is Required for Erythroblast Self-Renewal

(A) SREs and ESREs were isolated from restricted and extensive phases of self-renewal, respectively. One representative growth curve of four independent ESRE cultures is shown with timing of SRE and ESRE isolation boxed.

(B) Analysis of Affymetrix data sets revealed upregulation of genes associated with the PRC1 in ESREs and SREs compared to primary ProEs. Several known targets of Bmi-1 are significantly downregulated in ESREs/SREs compared to ProEs (mean ± SEM; N = 4 independent replicates for ESREs/SREs; N = 5 independent replicates for ProEs). p value was calculated using one-tailed Student’s t test. ∗p < 0.05; ∗∗p < 0.01.

(C) Bmi-1 transcripts are expressed at significantly higher levels in ESREs compared to primary CFU-Es, ProEs, and maturing erythroblasts (EryBs) (mean ± SEM; N = 3 independent replicates). p value was calculated using one-tailed Student’s t test. ∗p < 0.05.

See also Figure S1.

(D) shRNA-mediated knockdown of Bmi-1 rapidly decreased ESRE proliferation after puromycin selection (one representative culture of three independent experiments).

(E) PTC-209, a BMI-1 inhibitor, caused a dose-dependent inhibition of ESRE proliferation compared to vehicle (DMSO) control culture. One representative culture of six independent experiments is shown.