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. 2012 Dec 4;23(3):340–350. doi: 10.1038/cr.2012.164

Figure 4.

Figure 4

hnRNP I is a RoR binding partner. (A) A representative silver-stained gel picture showing a doublet band unique to E4-d2. Mass spectrometry analysis indicated that this band is hnRNP I. (B) Verification of the interaction of RoR with hnRNP I by RNA precipitation and western blot. Samples were prepared in the same way as in (A) and the membrane was probed with hnRNP I antibody. (C) RT-PCR detection of RoR after the hnRNP I immunoprecipitation using primers ROR-Exon4-RT-5.1A and ROR-Exon4-RT-3.1A (Supplementary information, Table S1). (D) Suppression of the doxo-induced p53 by hnRNP I-siRNAs. MCF-7 cells were first transfected with control siRNA or hnRNP-siRNAs (mixed pool), and then treated with 0.5 μg/ml doxo for 16 h before harvesting cells for western blot. (E) Effect of ectopic expression of hnRNP I on p53. MCF-7 cells were first transfected with vector or hnRNP I, and then treated with 0.2 μg/ml doxo for 16 h before western blot. The purpose of using a low concentration of doxo was to not allow p53 induction to reach the maximum so that we would be able to detect any further increase in the p53 level by hnRNP I.