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. 2007 Oct 12;8(11):1052–1060. doi: 10.1038/sj.embor.7401088

Figure 4.

Figure 4

RNA binding motif protein 4 is required for microRNA-guided gene silencing. (A) SiRNAs against the indicated proteins were pre-transfected into HeLa cells. After 2 days, a luciferase reporter containing a complementary binding site for miR-21 was transfected. (B) Experiments were carried out as described in (A). A luciferase reporter construct containing the 3′-UTR of KRAS was used (left panel). 2′OMe inhibitors of the indicated miRNAs were pre-transfected into HeLa cells (right panel). (C) Experiments were carried out as in (A). Luciferase reporter constructs carrying the 3′-UTRs of the indicated mRNAs were transfected and normalized to control siRNA. (D) Schematic illustration of RBM4. (E) Lysates from HEK 293 cells transfected with the indicated constructs were immunoprecipitated using Flag antibodies, and northern blotting against miR-19b was performed. GFP, green fluorescent protein; HEK, human embryonic kidney; KRAS, Kirsten rat sarcoma viral oncogene homologue; miRNA, microRNA; RBM4, RNA binding motif protein 4; siRNAs, small interfering RNAs; TNRC6B, trinucleotide repeat containing 6B; UTR, untranslated region.