Skip to main content
. Author manuscript; available in PMC: 2010 Mar 1.
Published in final edited form as: J Cell Physiol. 2009 Mar;218(3):593–602. doi: 10.1002/jcp.21632

Figure 6.

Figure 6

Oligomerization, glycosylation and different localization of soluble CRF1 isoforms in human HaCaT keratinocytes. Cells were transfected with plasmid DNA coding fusion of CRF1 isoforms with EGFP (a, c–h) or plasmids coding EGFP alone (N2). The EGFP tagged receptors was detected in cell lysates by Western Blotting (WB) with mouse anti-GFP antibody (1:500) and secondary anti-mouse antibody (1:4,000). A: High molecular weight complexes (HMCs) of isoforms: α c, d, f, and h. B: Dissociation of HMCs of CRF1h isoform; C, untreated control; 1, lysate treated with PNG-ase F, 2–6 M Guanidinium hydrochloride, 3–50 mM DTT. C: Detection of CRF1-EGFP fusion protein in membrane (M) or cytoplasm (C) fractions isolated from cells (C) or conditioned media (D, E). Extracellular localization of CRF1-EGFP fusion proteins detected by direct fluorescence of EGFP (D) or WB of proteins precipitated from media with TCA followed by detection with anti-GFP antibodies (mouse anti-GFP 1:500, anti-mouse 1:4,000) (E).