Fig. 8.

PMA and chelerythrine have no effect on the total expression or surface expression of hASIC1b. A and B: immunoblots showing hASIC1b expression with and without PMA (A) or chelerythrine (B) treatment in total membranes prepared as described in materials and methods 3 days postinjection of oocytes with RNA for WT, S40A, or S499A hASIC1b. U, uninjected oocytes. Equivalent amounts of protein (30 μg) were loaded on each lane and separated by SDS-PAGE. Proteins were transferred to PVDF membranes, which were blotted for hASIC1 (1:200) and actin (1:5,000) as a loading control. As shown by the actin immunoblot, these experiments were repeated 2–4 times. C: mean normalized luminescence of oocytes expressing hemagglutinin (HA)-tagged WT hASIC1b. There was no statistically significant difference between WT hASIC1b surface expression in control or treated oocytes (P = 0.628 by one-way ANOVA). Shown is a summary of 3 experiments. The number in parentheses on top of each bar indicates the number of individual oocytes measured.