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. 2016 Apr 4;7:11169. doi: 10.1038/ncomms11169

Figure 2. The anti-angiogenic effect of miR-192.

Figure 2

(a) Top 15 molecules downregulated by miR-192. Whole-genome microarray was performed at 48 h post transfection in SKOV3ip1 cells (n=3). (b) The effect of miR-192 on pro-angiogenic factors. (c) Transcription factors predicted to regulate >5 angiogenic factors and are potential direct targets of miR-192. The lines indicate potential interactions between the transcription factors and the pro-angiogenic genes. (d) The effect of miR-192 on the levels of transcription factors that can potentially mediate the broad anti-angiogenic function of miR-192. The mRNA levels of each transcription factor was assessed at 24 h post transfection in SKOV3ip1 cells (n=3, Student's t-test). (e) Relative (Rel.) luciferase activity normalized to empty control for EGR1 or HOXB9 3′-untranslated region (UTR). Mutated constructs have predicted miR-192 binding site deleted. SKOV3ip1 cells were transfected with miR-192 and UTR luciferase constructs using FuGene transfecting reagent, and luciferase assay was performed at 24 h post transfection (n=4, Student's t-test). (f) Tube formation potential was assessed in RF24 cells after exposing to conditioned media collected from SKOV3ip1 cells treated with siEGR1 and/or siHOXB9 (48 h post transfection, n=5, Student's t-test). (g) Effect of EGR1 or HOXB9 silencing on levels of angiogenic factors. SKOV3ip1 cells were treated with siRNAs and RNA was isolated at 48 h post transfection (n=3, Student's t-test). Scale bar, 100 μm. All bars and error bars represent mean values and the corresponding s.e.m. (*P<0.05; **P<0.01; ***P<0.001; and ****P<0.0001).