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. Author manuscript; available in PMC: 2011 Apr 13.
Published in final edited form as: Nat Neurosci. 2005 Feb 13;8(3):322–331. doi: 10.1038/nn1407

Figure 2.

Figure 2

CPG15 is predominantly expressed in a soluble secreted form. (a) Schematic of CPG15-FLAG-IRES-EGFP construct. The Flag peptide sequence (gray) was inserted between the secretion signal (SS) and the CPG15 core domain. Tagged, full-length CPG15 was then cloned upstream of an internal ribosome entry site (IRES) and EGFP. (b) HEK293T cells transfected with CPG15-FLAG-IRES-EGFP (green) were cocultured with untransfected cells (white) for 2 d. (c) Untransfected cells (outlined by square in b) show CPG15 immunoreactivity (white arrow). Scale bar, 10 μm. (d) HEK293T cells cocultured with cells transfected with a Flag-tagged control protein show no membrane staining. (e) Western blots of cell extracts and supernatants from CPG15-FLAG-IRES-EGFP transfected HEK293T cells either untreated or treated with phospholipase C (PLC). (f) Western blots of membrane and soluble fractions from E14 and E18 brains, and adult cortices, probed with antibody against CPG15. Staining for the transferrin receptor (Tf-rec.) and the enzyme Akt serve as membrane- and soluble-fraction controls, respectively. Ponceau staining (Pon.) serves as a loading control.