Skip to main content
. 2010 Sep 23;5(9):e12964. doi: 10.1371/journal.pone.0012964

Figure 4. GPR54 positively regulates ERK1/2 activation in a β-arrestin-2-dependent manner.

Figure 4

Representative autoradiograph (A) and densitometric analysis (B) showing the expression of total and activated ERK1/2 in GPR54 overexpressing β-Arr2 KO and corresponding wild type parental (WT2) MEFs following stimulation with 100 nM Kp-10 (for the indicated time points). (C) Representative Western blot confirming absence and presence of expression of FLAG-GPR54 in non-electroporated (NT) and FLAG-GPR54 overexpressing β-Arr2 KO and WT2 MEFs, respectively. (D) Representative western blot showing the expression of total and activated ERK 1/2 following treatment of the β-Arr2 KO and corresponding wild type parental (WT2) MEFs with 10 ng/ml EGF (for the indicated time point). This was used as a control to assess whether the ERK1/2 pathway was still functional in these cells. Representative autoradiograph (E) and densitometric analysis (F) showing the expression of total and activated ERK1/2 following stimulation with 100 nM Kp-10 (for indicated time points) of FLAG-GPR54 overexpressing β-Arr2 KO MEFs co-transfected with either GFP vector (grey bars) or β-Arr2-GFP (white bars). All western blot analyses were done using monoclonal anti-ERK1/2 and anti-phospho ERK1/2 antibodies. Monoclonal anti-β-Arr2 and anti-DDK (FLAG) antibodies were also used. The data represent the mean ± S. E. of 4 independent experiments (B) or the mean ± S. E. of 3 independent experiments (F). ## P<0.01 vs 0 min. control (within the specific cell line). *P<0.05, ** P<0.01vs respective wild-type (or ‘add-back’) control at the indicated time point.