Figure 5. GLO1 knockdown reduces ALDH1^high cell numbers and induces apoptosis.

(A) Immunoblot analysis of GLO1 silenced in MCF 10A, MDA-MB 157 and MDA-MB 468 using GLO1-targeted siRNAs. β-actin was used as an internal control. (B) GLO1 activity measured 48 h after transfection of targeted siRNA in MCF 10A, MDA-MB 157 and MDA-MB 468 cells (1.0 × 10⁴ cells/well). GlO1 activity is expressed in arbitrary units (A.U.). ^*p < 0.05, ^**p < 0.01; Tukey's test. All the experiments were performed in duplicate. Data represent the mean ± SE (three independent experiments). (C) Numbers of ALDH1^high cells isolated from MDA-MB 157 cells and MDA-MB 468 cells after GLO1 knockdown were measured in ALDEFLUOR assays. ^*p < 0.05; Students t-test. Data represent the mean ± SD (MDA-MB 157: three independent experiments, MDA-MB 468: four independent experiments). (D) Numbers of active caspase-3-positive ALDH1^high cells isolated from MDA-MB 157 and MDA-MB 468 cells after with GLO1 knockdown. ^*p < 0.05; Students t-test. Data represent the mean ± SD (MDA-MB 157: three independent experiments, MDA-MB 468: four independent experiments). (E) Trypan blue staining for ALDH1^high cells isolated from MDA-MB 157 and MDA-MB 468 cells after GLO1 knockdown. ^*p < 0.05; Students t-test. Data represent the mean ± SD (three independent experiments).