Figures & data
Figure 1. LncRNA NNT-AS1 expression is elevated in ccRCC tissues and cell lines. A Schematic representation of NNT-AS1. B NNT-AS1 expression level in ccRCC tissues and matched normal tissues by qRT-PCR. C NNT-AS1 expression level in ccRCC cell lines and normal renal cell line HK-2 by qRT-PCR. ***P < 0.001
Figure 2. Silencing of NNT-AS1 restrains ccRCC proliferation and metastasis. A Verification of NNT-AS1 siRNA efficiency by qRT-PCR analysis. B Effect of NNT-AS1 knockdown on ccRCC cells proliferation. C and D Effect of NNT-AS1 knockdown on ccRCC metastasis. E Effect of NNT-AS1 knockdown in in vivo xenografted animal model. *P < 0.05, ** P < 0.01, ***P < 0.001
Figure 3. Overexpression of NNT-AS1 promotes ccRCC proliferation and metastasis. A Verification of the NNT-AS1 plasmids overexpression efficiency by qRT-PCR analysis. B Effect of NNT-AS1 overexpression on ccRCC cells proliferation. C and D Effect of NNT-AS1 overexpression on ccRCC metastasis. ** P < 0.01, ***P < 0.001
Figure 4. NNT-AS1 can directly bind to miR-137. A qRT-PCR analysis of nuclear and cytoplasmic RNA fraction in ccRCC cells. B AGO-2 RNA-IP assay followed qRT-PCR analysis showing binding capacity between NNT-AS1 and AGO2. C Left, the predicted binding site of WT NNT-AS1 or MUT NNT-AS1 with miR-137; right, luciferase activity was detected in HEK-293 T cells. D and E Effect of the NNT-AS1 knockdown or overexpression on miR-137 expression. F Correlation analysis between NNT-AS1 and miR-137. *P < 0.05, ** P < 0.01, ***P < 0.001
Figure 5. NNT-AS1 activity is partially mediated through negatively regulation of miR-137. A miR-137 expression level in ccRCC tissues and matched normal tissues. B Verification of the effect of miR-137 mimics or inhibitors by qRT-PCR analysis. C Effect of siNNT-AS1 or siNNT-AS1 plus miR-137 inhibitors on ccRCC cells proliferation. D and E Effect of siNNT-AS1 or siNNT-AS1 plus miR-137 inhibitors on ccRCC cells metastasis. *P < 0.05, ** P < 0.01, ***P < 0.001
Figure 6. NNT-AS1 indirectly regulates YBX-1 expression via competitively binding with miR-137. A Left, the predicted binding site of miR-137 with wild type 3ʹ UTR of YBX-1 or mutant YBX-1; right, luciferase activity was detected in HEK-293 T cells. B miR-137 expression level in ccRCC tissues and matched normal tissues. C Correlation analysis showed a negatively correlation between YBX-1 and miR-137. D Effect of miR-137 mimic on YBX-1 expression. E effect of miR-137 inhibitors on YBX-1 expression. E Effect of siNNT-AS1 and siNNT-AS1 plus miR-137 inhibitors on YBX-1 expression. G Effect of siNNT-AS1 and siNNT-AS1 plus YBX-1 overexpression on ccRCC cells proliferation. H Effect of siNNT-AS1 and siNNT-AS1 plus YBX-1 overexpression on ccRCC cells metastasis. *P < 0.05, ** P < 0.01, ***P < 0.001
