Fig. 4: ZC3H12D recognizes the specific sequence of IL1β-mRNA.
From: Extracellular mRNA transported to the nucleus exerts translation-independent function
a Scheme to determine ZC3H12D binding site in mIL1β-mRNA (NM_008361) 3′-UTR. Each number denotes nucleotide position. b EMSA. mIL1β probes (EMSA probes 1–9; see Supplementary Fig. S4a for sequences. Nucleotide positions of each probe are displayed at the top) were assayed with or without ZC3H12D protein. Probes 1–9 are 50 nt long, and the 3′-end is biotin-labeled. Arrow indicates the band shift due to the binding of hZC3H12D on probe 5. c EMSA competition assay. ZC3H12D protein and biotin-labeled probe 5 (50 nM) were mixed with a 100-fold excess amount of non-labeled probes 1–9 (5 μM). d EMSA competition assay. ZC3H12D protein and biotin-labeled probe 5 (50 nM) were mixed with a 100-fold excess amount of the non-labeled probes 5-1–5-7 (5 μM). The competitors are 20 nt long and part of probe 5 (Supplementary Fig. S4b). The graph indicates the relative intensity of the top band [= top band / (top band + bottom band) × 100]. The top band position is marked with an arrow. Experiments were repeated twice with similar results. Source data are provided as a Source Data file.
