Fig. 3

R-RXRE is important for pre-miR-103a-2 binding to RXRα. a Schematic diagram of pre-miR-103a-2 wild-type (WT) and mutants (M1–M5). The R-RXRE is underlined and in black, and the sequences of the mature miRNAs are in magenta. The substituting bases are shown in bold capital letters. b In vitro transcribed pre-miR-103a-2 and its mutants were pulled down by the recombinant GST-RXRα/DBD or GST protein followed by quantifying by qRT-PCR. The PCR values of GST-RXRα/DBD fusion protein was normalized to that of GST protein. c RNA-IP analysis of the association of Flag-RXRα with pre-miR-103a-2 and its mutants in Flag-RXRα stable AD293 cells. d HEK293T cells were transfected with pGL6-TA-RXRE-luciferase and Myc-RXRα expression plasmids together with pre-miR-103a-2 or its mutant expression plasmids for 24 h. Cells were then treated with or without 9-cis-RA (10^–7 M) for 12 h. The value of the firefly luciferase activity was normalized by the renilla luciferase activity to generate the relative luciferase (Luc) activity. Statistical significance of the difference between the second group and the other groups are shown. Data are presented as mean ± SEM (n = 3), ****P < 0.0001, and ns means not significant