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. 2016 Jul 4;16:379. doi: 10.1186/s12885-016-2470-3

Fig. 4.

Fig. 4

SIAH1 and PTP41A genes are miR-944 targets. (a) Schematic representation indicating the 3´UTR sequence of PRKCA, PTP4A1 and SIAH1 genes cloned in pmiR-report vector. Boxes indicate the miR-944 binding sites in target genes. (b, c, d) Luciferase reporter assays. MDA-MB-231 cells were co-transfected with miR-944 (or scramble as control) and pmiR-LUC-PRKCA-3´UTR (b), pmiR-LUC-PTP4A1-3´UTR (c) or pmiR-LUC-SIAH1-3´UTR (d) plasmids and relative luciferase activity was measured as described in methods. Results are expressed in light units. Bars represent the mean of three independent experiments performed three times ± S.D. (e) Immunodetection of SIAH1 by Western blot assays in MDA-MB-231 cells. Lane 1, MDA-MB-231 control cells; lane 2, MDA-MB-231 cells transfected with miR-944. (f) Immunodetection of SIAH1 in breast tumors and normal mammary tissues. β-tubulin was used as internal control. (g) Densitometry analysis of immunodetected bands in F. Pixels corresponding to β-tubulin were used to normalize SIAH1 expression. NS, non- significant. *p < 0.05; **p < 0.01; ***p < 0.001