Supporting information for Tanka et al. (2003) Proc. Natl. Acad. Sci. USA, 10.1073/pnas.0837789100

Supporting Figure 9

Fig. 9.

 Northern blot analysis of mouse neuropeptide B (NPB) and neuropeptide W (NPW) mRNA. Northern blot with 7 μg of poly(A)-rich RNA per lane from various mouse tissues were rehybridized with designated probes. High levels of NPB mRNA expression were observed in the stomach, spinal cord, and testis, whereas the strongest NPW mRNA expression was detected in the lung. A weak NPB mRNA expression was also observed in the brain, liver, and kidney. NPW mRNA was also detected in the stomach. Northern blots for human tissues yielded the expression pattern of the NPB, NPW, and GPR7 mRNAs similar to the data described in another paper (ref. 1; data not shown). Total RNA of mouse tissues was extracted with the RNA STAT-60 reagent (TEL-TEST, Friendswood, TX), and Poly(A)-rich RNA prepared with the Oligotex mRNA mini kit (Qiagen, Valencia, CA). Northern blot was done as described (2), with a 0.4-kb StyI–NdeI fragment of the mouse NPB cDNA and a 0.47-kb EcoRI–HindIII fragment of the rat NPW cDNA as probes.

1. Brezillon, S., Lannoy, V., Franssen, J. D., Le Poul, E., Dupriez, V., Lucchetti, J., Detheux, M. & Parmentier, M. (2003) J. Biol. Chem. 278, 776–783.

2. Xiong, Y., Tanaka, H., Richardson, J. A., Williams, S. C., Slaughter, C. A., Nakamura, M., Chen, J. L. & Yanagisawa, M. (2001) J. Biol. Chem. 276, 28471–28477.