Figure 3: TUG1 antagonizes miR-145 and maintains expression of stemness-associated genes.
From: Targeting the Notch-regulated non-coding RNA TUG1 for glioma treatment

(a) Heatmap shows commonly upregulated miRNAs upon inhibition of TUG1 in GSCs (1228 and 222). Colour corresponds to expression level as indicated in the log2-transformed scale bar below the matrix. Red and blue reflect high and low levels, respectively. (b) Effect of TUG1 depletion on miR-145 expression in GSCs (1228 and 222). y-axis indicates relative miR-145 expression level compared with that in si-NC-treated cells. Expression levels were normalized to internal RNU6B. *P<0.01, Student’s t-test. (c) RNA-FISH analysis of TUG1 (red) and miR-145 (green) in GSCs treated with si-NC or si-TUG1. Nuclei are stained with DAPI. Scale bars, 10 μm. (d) The spot numbers relating to TUG1 and miR-145 detection were quantified per cell in GSCs treated with either si-NC or si-TUG1. *P<0.001, Student’s t-test. (e) Schematic of mutated (TUG1 MUT) or deleted (TUG1 DEL) TUG1. (f) Effect of exogenous miR-145 on TUG1, SOX2 and MYC expression in GSC-pE-Nes-222. Partial TUG1 transcripts (TUG1 WT, TUG1 MUT and TUG1 DEL as shown in (e) were added to GSC-pE-Nes-222 treated with the precursor molecule of miR-145 (miR-145). Expression levels of endogenous TUG1, SOX2 and MYC were measured. Values are indicated relative to abundance of negative control miRNA precursor (NC) treated cells. *P<0.01, Kruskal–Wallis analysis. (g) Phase-contrast and Nestin-EGFP images of miR-145+ TUG1 WT, +TUG1 MUT or +TUG1 DEL cells as shown in f. Scale bars, 100 μm. (h) Intensity of Nestin-EGFP (left) and number of viable cells (right) were quantified. Viable cells were assessed by trypan blue staining. *P<0.01, Kruskal–Wallis test. For all the experimental data, error bars indicate s.d. (n=3).