Total RNA was extracted from NTera-2 cells using the Qiagen RNeasy Mini kit following the manufacturer's protocol (Qiagen, Valencia, CA, USA). The target RNA was amplified by a one-step RT-PCR kit (GMbiolab, Taichung, Taiwan). Forward and reverse primers were as follows: GFAP (forward), 59-GTGGGCAGGTGG-GAGCTTGATTCT-39 and (reverse), 59-CTGGGGCGGCCTGG-TATGACA-39 [27] ; and internal control b-actin (forward), 59-TGGAATCCTGTGGCATCCATGAAAC-39 and (reverse), 59-TAAAACGCAGCTCAGTAACAGTCCG-39. The RT-PCR consisted of two programs. First was complementary cDNA synthesis: one cycle at 50uC for 30 min and one cycle at 94uC for 2 min. The other was second-strand cDNA synthesis and the PCR consisted of 35 cycles at 94uC for 30 s, 59uC for 30 s, and 72uC for 45 s; followed by a final extension step at 72uC for 10 min.