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. Author manuscript; available in PMC: 2006 Oct 1.
Published in final edited form as: Am J Physiol Cell Physiol. 2005 May 18;289(4):C934–C945. doi: 10.1152/ajpcell.00092.2005

Figure 2. OGD-mediated stimulation of NHE1+/+ activity is attenuated by PD98059.

Figure 2

Following either normoxia or OGD/REOX in NHE1+/+ astrocytes, a NH3/NH4+ prepulse was applied and JH+ determined. In the PD98059 studies, 30 μM PD98059 was present during OGD and REOX. A. Representative pHi response in single cells to a prepulse following either normoxia (2.25 h), OGD (2 h)/REOX (15 min), or OGD/REOX in the presence of PD98059. B-D. During recovery from a prepulse, JH+ was determined and plotted. Data were fitted with a sigmoid curve to determine Km and Vmax. E. Following OGD or OGD/REOX, NHE1+/+ astrocytes were lysed and lysate was separated by 12% SDS-PAGE, and transferred to nitrocellulose membrane. The blot was probed with either anti-ERK antibody (ERK1/2) or anti-phospho-ERK1/2 antibody (p-ERK1/2). Representative images of three experiments is presented. F. Immunostaining for GFAP (a, c) and p-ERK1/2 (arrows, b, d) in NHE1+/+ astrocytes under normoxic control (a, b) or 1 h OGD (c, d). Scale bar: 120 μm. Representative image of three experiments is presented. Inset in F(b): image taken when anti-p-ERK1/2 antibody was omitted.