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. Author manuscript; available in PMC: 2013 Feb 24.
Published in final edited form as: Exp Cell Res. 2012 May 29;318(16):2004–2013. doi: 10.1016/j.yexcr.2012.04.015

Fig. 1.

Fig. 1

ERK phosphorylation induced by OBDL in OX2R-CHO cells. (A) Immunofluorescence staining of OX2R in CHO-OX2R cells. OX2R in methanol fixed CHO cells was stained with goat anti-OX2R antibody (SC-8074 Santa Cruz Biotech) and DNA was counterstained with DAPI. (B) Comparison of ERK phosphorylation induced by orexin B and OBDL. Time courses of ERK phosphorylation induced by 500 nM of orexin B and OBDL were used to compare the function difference of orexin B and OBDL. (C) Orexin B dose-dependent phosphorylation of ERK by Western blot and FACE assay. ERK phosphorylation was detected by anti-p-ERK and –ERK antibodies (9101 and 9102, Cell Signaling) at 1:1000 dilution. ERK phosphorylation at each orexin B concentration in FACE is the mean value of four repeat measurements. (D) Time course of ERK phosphorylation induced by orexin B. OBDL (500 nM) was used to treat cells for different times. p-ERK was normalized by total detected ERK. (E) Time course of ERK phosphorylation induced by orexin A. 500 nM of orexin A was used to treat CHO-OX2R cells for up to 40min. p-ERK was normalized by total ERK. The mean and P-value were calculated from three independent repeated experiments. Significance of p-value is less than 0.05.