Skip to main content
. Author manuscript; available in PMC: 2018 Dec 1.
Published in final edited form as: Cell Signal. 2017 Sep 28;40:222–229. doi: 10.1016/j.cellsig.2017.09.021

Figure 4. Phos-tag analysis indicates that FGF2 stimulates GC-B dephosphorylation.

Figure 4

RCS cells, or RCS cells virally transduced with wild type GC-B (RCS-GC-B) were treated with 1 μg/ml heparin in the absence or presence of 100 ng/ml FGF for 20 min. Cells were lysed and crude membranes were immunoprecipitated with an antibody against the C-terminus of GC-B. The immunocomplexes were washed and fractionated on a Phos-tag gel and western blotted with an antibody against the extracellular domain of GC-B. (A–C) RCS cells that endogenously express GC-B. Note that standard molecular weight markers do not accurately reflect MW on Phos-tag gels and were thus not included. (B) Quantitation of densitometry results as shown in (A) and described in the text; results are from six independent determinations from two experiments (C) GC assays of the same six sets of membranes used for (B) under CNP-stimulated conditions. (D–F) Samples from RCS cells virally transduced with WT-GC-B. (E) Quantitation of densitometry results as shown in (D); results from five independent determinations from two blots (F) GC assays of the same five sets of crude membranes used for (E) under CNP-stimulated conditions. **, ***, and **** indicates significance vs. control at p<0.01, 0.001, and 0.0001, respectively, as determined by unpaired, two-tailed student’s t-test.