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. Author manuscript; available in PMC: 2019 May 20.
Published in final edited form as: Sci Signal. 2018 Nov 20;11(557):eaau7632. doi: 10.1126/scisignal.aau7632

Fig. 6. miR-27a over-expression silences PAX3:FOXO1.

Fig. 6.

(A and B) Western blot of PAX3:FOXO1 protein abundance in Rh30 and CF-1 cells transfected with mimics of miR-27a (10 μM for 72 hours) or a negative control (ctrl). Also shown is blotting in lysates from untransfected control cells (unt). Blots are representative of N=3 independent experiments. (C and D) Light microscopy images of Rh30 and CF-1 cells transfected with mimics of miR-27a. Images are representative of N=3 independent experiments. Scale bar, 50 μM. (E to H) qPCR analysis of miR-27a (E and F) or miR-485p (G and H) expression in Rh30 and CF-1 cells treated for 72 hours with ENT (1 μM) or PAN (45 nM). (I and J) qPCR of miR-27a expression in Rh30 and CF-1 cells transfected with HDAC2- or HDAC3-targeted siRNA (100 nM). Data were normalized to U6snRNA expression. Gene expression was quantified using the 2^-dCt method. Data are means ± SD, N=3 independent experiments each in triplicate.; * P < 0.05, **P < 0.01, and ***P < 0.001 by a two-sided Student’s t tests. (K) Summary of the HDAC3–SMARCA4–miR-27a–PAX3:FOXO1 regulatory circuit targeted by ENT.