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. 2018 Sep 28;11:6339–6350. doi: 10.2147/OTT.S178001

Figure 4.

Figure 4

Insulin receptor substrate 1 (IRS1) is a direct target of miR-1225-5p in glioblastoma.

Notes: (A) Protein expression in U87 and U87-1225 cells was measured by Western blotting. (B) The expression of IRS1 in U87 and U87-1225 (stably expressing miR-1225-5p) cells was measured by reverse transcription PCR (RT-PCR) and normalized against U6. (C) Top, wild-type (wt) miR-1225-5p target sequences of IRS1 3′-untranslated region (UTR) and mutant (mut) miR-1225-5p target sequences of IRS1 3′-UTR. Bottom, relative luciferase activity of wt or mut pGL3-IRS1-3′-UTR in U87 and U87-1225 cells. (D) The mRNA (top) and protein (bottom) expression level of IRS1 in glioblastoma cell lines (A172, SW1783, U87, LN-229, SW1088, and T98G) and normal human astrocytes was measured by Western blotting or RT-PCR. (E) Scatter plot of the RNA Sequence data (TCGA) of 63 samples from glioblastoma patients. The red line indicates an inverse correlation of expression between the samples for miR-1225-5p and IRS1 genes (R=−0.5132; P=2.248e-05). **P<0.01.