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. Author manuscript; available in PMC: 2010 Oct 5.
Published in final edited form as: Nature. 2009 Dec 13;463(7279):364–368. doi: 10.1038/nature08697

Figure 1. hnRNP proteins bind specifically to sequences flanking E9.

Figure 1

a, Schematic diagram of PKM splicing. b, Position of probes spanning the E9 or E10 5′ splice sites (top). After ultraviolet crosslinking, proteins were detected by autoradiography (bottom). Position of molecular mass standards in kDa is indicated at left. c, Affinity chromatography using EI9(50–68). Bound proteins were separated by SDS–PAGE and Coomassie stained. Bands excised for mass spectrometry are indicated. d, Sequence of EI9(50–68); the putative hnRNPA1/A2 binding site is indicated in bold italics (top). Ultraviolet crosslinking with wild-type RNA, or RNA with a mutation in the putative hnRNPA1/A2 binding site, is shown in the bottom panel. e, Position of I8 and I9 (top). Ultraviolet crosslinking using I8 or I9 substrates is shown in the bottom left panel. Ultraviolet crosslinking reactions were immunoprecipitated with either anti-PTB (BB7) or anti-HA antibodies (bottom right panel). f, Ultraviolet crosslinking with I8 and the mutant derivative I8mu, sequences indicated above. Putative PTB binding sites in I8 are underlined.

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