β-Arrestin1/miR-326 Transcription Unit Is Epigenetically Regulated in Neural Stem Cells Where It Controls Stemness and Growth Arrest

Figure 4

βarr-1 overexpression impairs NSCs clonogenicity and proliferation. (a) Representative image of immunofluorescence staining of NSCs after overexpression of HA-βarr-1. Nuclei are counterstained with Hoechst. Scale bar: 5 μm. (b) WB analysis of endogenous stemness markers in NSCs after overexpression of HA-βarr-1. Loading control: actin. Representative Western blot images from 3 independent experiments. (c) mRNA expression levels of stemness markers in NSCs after overexpression of HA-βarr-1. (d) Oncosphere forming assay (number of colonies, left panel) and bromodeoxyuridine (BrdU) uptake (right panel) in NSCs after ectopic expression of HA-βarr-1. (e) Cell viability (MTT assay) in NSC after ectopic expression of HA-βarr-1. (f) WB analysis of cleaved Parp (Parp-c) in NSCs after overexpression of HA-βarr-1. Loading control: Gapdh. Representative Western blot images from 3 independent experiments. ((c), (d), and (e)) Data are means ± SD from 3 independent experiments. .

(a)

(b)

(c)

(d)

(e)

(f)