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. 2019 Jan 9;8(1):5. doi: 10.1038/s41389-018-0113-z

Fig. 3. miR590-5p regulation of FAM171A1 in breast cancer cell lines.

Fig. 3

a Chromatin Immunoprecipitation analysis showing lack of ERα recruitment onto the FAM171A1 promoter; pS2 and Cyclin D as positive controls. b Diagram showing three seed sequences at designated locations within the 3ʹ-UTR of the FAM171A1 gene. c Endogenous expression of miR590-5p in breast cancer cell lines as analyzed by qRT-PCR using TaqMan chemistry; ***p < 0.0003. d qRT-PCR showing the induction of preMIR590 expression upon ERα transfection in MDAMB231 cell line. The data were normalized to U6 snRNA. e Schematic diagram showing the putative ERα-binding region in the MIR590 promoter. Exon in light gray color, ERE and STAT5 binding sites in black color. f CHIP analysis of ERα binding onto the MIR590 promoter in MCF-7 and T47D cells. SG6K and DNAMT3A primers were used as negative controls. g pSi-CHECK2 and pRIP vector constructs showing the regions for cloning of 3ʹ-UTR region and MIR590 gene, respectively. h Effect of ERα cotransfection upon miR-590 promoter-luc reporter activity; *p-value < 0.05 (Student’s t-test). i Effect of miR-590-pRIP expression on the 3ʹ-UTR-pSi-CHECK2-luc activity; *p-value < 0.05 (Student’s t-test). Experiments (a, c, f, h, i) were performed in three biological replicates and d was performed two times each time with three technical replicates