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. 2014 Nov 5;111(46):16502–16507. doi: 10.1073/pnas.1419349111

Fig. 2.

Fig. 2.

Biochemical and proteomic detection of Oxtrs in osteoblast nuclei. MC3T3.E1 osteoblasts and Oxtr−/− primary osteoblasts (OBs) (negative control) were treated with Oxt (10−8 M) for 20 and 40 min. (A) Western blots show Oxtrs in the cytosolic and nuclear fractions at 40 min. Markers included t-Erk (cytosol), Rab11 (perinuclear), and Hist1h1 (nuclear). Proteins in the immunoprecipitate (with anti-Oxtr antibody) were identified using MS-Fit software (http://prospector.ucsf.edu/prospector/cgi-bin/msform.cgi?form=msfitstandard), and all other proteins were recognized as nuclear proteins. (B) Analysis of the spectra (FindPept database) revealed five peptides corresponding to Oxtr intracellular loops in the in-gel band (38 kDa) at m/z of 1,851.98 [(R)/TTRHKHSRLFFFMK/(H); residues 66–79], 1,059.61 [(R)/SLRRRTDR/(L); residues 146–153], 3,410.05 [(R)/LKTAAAAAAAEGSDAAGGAGRAALARVSSVKLISKAK/(I); residues 232–268], 1,256.84 [(K)/LISKAKIRTVK/(M); residues 263–273], and 2,325.21 [(K)/GSRPGETSISKKSNSSTFVLSR/(R); residues 353–374]. ’, minute.