Fig. 3
From: Heme enables proper positioning of Drosha and DGCR8 on primary microRNAs
Heme enables DGCR8 to guide Drosha to the correct junction. a 5′ end-labeled in vitro processing assays of pri-miR-21, using MPHeme (130 nM), MPC352S (130 nM), MPΔHBR (260 nM), and MPCTT (1.04 μM). Correct (black asterisk) and incorrect (red asterisk) products are indicated. DGCR8ΔHBR lacks the HBR but contains the tandem dsRBDs and C-terminal tail. Deletion of HBR and dsRBDs yields the C-terminal tail (DGCR8CTT). b, c EMSAs comparing binding affinities of MPΔHBR to basal b and apical c junctions of pri-miR-21. Schematic diagrams of the RNA are shown next to free RNA bands (arrow), with mature sequence shown in light blue. The basal junction RNAs are annealed RNA duplexes lacking the terminal loop. The apical junction RNAs are the pre-miR fragments. Protein concentrations are (left to right): 3.33, 6.67, 10, 13.32, and 16.67 nM. d 5′ end-labeled in vitro processing assays of pri-miR-125a, similar to a. e, f EMSAs similar to b and c, for pri-miR-125a
