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. 2020 Oct 30;21(21):8132. doi: 10.3390/ijms21218132

Figure 6.

Figure 6

Manipulating SF-miR-7704 levels. (A–C) Inhibition of miR-7704 expression upregulates the expression of HAGLR. Results of quantitative real time PCR analysis of the effect of the inhibition of miR-7704 on the nuclear expression of HAGLR in MCF-10A (A), MCF-7 (B), and MDA-MB-231 cells (C) are shown. Transfection with Anti-miR-7704 inhibitor resulted in the down-regulation of miR-7704 (upper panel), and an increase in the expression level of HAGLR mRNA (lower panel) relative to the control. For the control we used the non-silencing Anti-miR (see Materials and Methods). The average of 3 independent biological preparations are shown, including the standard error. The expression levels of HAGLR was normalized to the internal control of ß-actin expression from the same preparation. (D) The FACS analyses are based on staining cells with Hoechst to quantify the DNA content following transfecting the MDA-MB-231 cells with miR-7704 or with its empty vector. The sorting is based on co-expression of GFP (488 nm; positive cells). The results from the overexpression with the empty vector (top) and the of same vector with miR-7704 (bottom) are shown. The fraction of the cells which accounts for an excess of DNA (>2 n; S/G2/M in cell cycle) are marked by a red frame.