Figure 3: TUG1 antagonizes miR-145 and maintains expression of stemness-associated genes. | Nature Communications

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

Figure 3

(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).

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