Figure 3. Bloodlinc Modulates the Core Gene Program of Terminal Erythropoiesis.

(A) Left: expression change (log₂ fold change over control) of 488 genes differentially and reciprocally regulated upon Bloodlinc shRNA-mediated inhibition or ectopic overexpression (OV) after 24 hr of ex vivo differentiation. Right: top non-redundant gene ontology (GO) biological processes enriched among reciprocally regulated genes.

(B) Bloodlinc modulation impacts erythropoiesis-repressed (top) and erythropoiesis-induced genes (bottom). Expression changes for select genes (left) during normal differentiation is shown as the log₁₀ expression ratio between BFU-Es and TER119⁺ erythroblasts (ERY). Their expression changes upon Bloodlinc depletion or overexpression (right) are shown as the log₂ expression ratio between treated versus control cells. Depletion data are pooled from two separate shRNAs.

(C) Erythropoiesis-induced genes are Bloodlinc-inducible. Gene set enrichment analysis of genes induced in erythroblasts versus progenitors (Alvarez-Dominguez et al., 2014) in Bloodlinc-depleted versus Bloodlinc-overexpressing cells. NES, normalized enrichment score.

(D and E) qPCR detection of erythroid markers reciprocally regulated in Bloodlinc-depleted (D) versus Bloodlinc-overexpressing (E) cells. Data are mean ± SEM from n = 2 experiments with n = 3 replicate measurements each. *p < 0.05 relative to control, t test.

See also Figure S5 and Table S2.