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. 2022 Oct 20;298(12):102630. doi: 10.1016/j.jbc.2022.102630

Figure 2.

Figure 2

HOTAIR is required for the self-renewal maintenance of breast CSCs.A, HOTAIR was silenced in breast CSCs by two independent shRNAs and analyzed by RT-PCR. HOTAIR-silenced stable cell lines were established. Data are shown as means ± SD. B, ALDH1+ (CSC) subpopulations were detected in HOTAIR-depleted cells by FACS analysis. Results are shown as means ± SD. C, HOTAIR depletion causes a diminished oncosphere-forming capacity in MCF7 CSC. Cells were magnified by 100 folds and the scale bar represented 10 μm. D, different numbers of HOTAIR-silenced or control breast CSC of MCF7 were diluted and subcutaneously implanted into NOD/SCID nude mice. Tumor-free mice were measured 1 month after injection. N = 10 for each group. E, different numbers of HOTAIR-silenced or control breast CSC of MDA-MB-453 were diluted and subcutaneously implanted into NOD/SCID nude mice. Tumor-free mice were measured 1 month after injection. N = 10 for each group. F, HOTAIR-silenced or control breast CSC of MCF7 were diluted and subcutaneously implanted into NOD/SCID nude mice. Tumors were observed over 1 month. N = 10 for each group. G, HOTAIR-silenced or control breast CSC of MDA-MB-453 were diluted and subcutaneously implanted into NOD/SCID nude mice. Tumors were observed over 1 month. N = 10 for each group. H, pluripotent transcription factors were analyzed in HOTAIR-depleted cells by quantitative real-time PCR. Results are shown as means ± SD in MDA-MB-453. I, the level of pluripotent transcription factors were analyzed in HOTAIR-depleted cell by Western blotting. β-actin was used as a control. CSC, cancer stem cell; HOTAIR, Hox transcript antisense intergenic RNA (∗ indicates p < 0.05; ∗∗ indicates p < 0.01; ∗∗∗ indicates p < 0.001.).