NRG1 induced an increase of EAAC1 expression through activation of the ErbB4 receptor.
A, Western blot analysis of EAAC1 (upper panel), ErbB2 (middle panel), and ErbB4 expression (lower panel) in C6 cells. The cell lysates were loaded with a different amount of protein (10, 20, 30, and 40 μg). B, inhibition of NRG1 by AG1478 induced the increase of EAAC1 expression. C6 cells were pretreated without or with 5 μm AG879 (ErbB2 inhibitor) and 5 μm AG1478 (ErbB4 inhibitor) for 15 min prior to the addition of NRG1 (5 nm, final concentration) for 30 min. C, quantification analysis of data in B. n = 6. *, p < 0.05. D, inhibition of ErbB4 blocks NRG1-induced increases of EAAC1 in primary cortical neurons. E, bar graph summarizes the data in D. n = 6. *, p < 0.05. F, no effect of NRG1 was observed on mRNA levels of EAAC1 in C6 cells (n = 6). G, qRT-PCR was performed to examine EAAC1 expression in cultured primary hippocampal neurons after treatment with 5 nm NRG1.