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. 2013 May 1;27(9):991–996. doi: 10.1101/gad.211243.112

Figure 3.

Figure 3.

DHX36 interacts with the pre-miR-134 terminal loop. (A) Scheme of the purification strategy used to isolate pre-miR-134-interacting proteins from P15 rat brain extract. (B) Coomassie gel of pull-downs using rat P15 brain extract and the indicated pre-miRNAs. Arrows point to bands that were identified by mass spectrometry as DHX9 (∼140 kDa) and DHX36 (∼110 kDa). (C) Western blot against the indicated proteins with extracts from pull-downs shown in B. (D) Western blot against DHX36 with extracts from the indicated pre-miRNA pull-downs. (E) Quantification of DHX36 binding in pre-miRNA pull-down experiments (pre-miR-134 = 1; mean ± SD, n = 3). (F) In vitro pre-miRNA cleavage assay. pre-miRNAs were incubated with recombinant Dicer for the indicated time in either the presence or absence of GST-DHX36. The position of pre-miRNAs and mature miRNAs is indicated. (G) Ratio of mature miRNA to pre-miRNA as an index for Dicer cleavage activity (mean ± SD, n = 4–5; conditions without DHX36 were set to 1 for both time points).