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. 2015 Oct 16;6(35):37852–37870. doi: 10.18632/oncotarget.5635

Figure 1. LincRNA-p21 represses stem-like properties and tumorigenicity of ALDH+ CSCs in vitro.

Figure 1

A. The levels of lincRNA-p21 were quantified by qPCR in ALDH+ CSCs and ALDH− non-CSCs, as well as human immortalized normal colon epithelial cells (NCM460) and normal liver cells (L-02). U6 was used as an endogenous control. CRC 1# and 2# represent two cases of primary CRC cells. B. Flow cytometric analysis of ALDH+ population in ALDH+ CSCs that were transduced with Lv-lincRNA-p21 (Lv-lnc-p21) or control lentiviral vector (Lv-ctrl) at 10 MOI for 7 days. C, D. ALDH+ CSCs were infected with Lv-lnc-p21 or Lv-ctrl (10 MOI). The diameter (C) and numbers of colonospheres (D) originating from single ALDH+ cells were measured with ImageJ software on Day 14. E. The numbers of soft agar colonies formed by these cells were counted and shown as means ± SD. Colonies with a diameter higher than 75 μm were counted. F. Stem cell markers, EpCAM, CD44, Lgr5, Nanog and Oct4, and differentiation markers Muc2 and CK-20 were examined by immunoblot analysis. Tubulin was a loading control. Representative graphs (B) or images (C, E, F) are shown. Data are presented as the mean ± SD (A, C, D, E) of each group from triple replicates. *P < 0.05, **P < 0.01.