mRNA was isolated using the RNeasy Mini kit and RNase-Free DNase set (Qiagen). Template cDNA was prepared by reverse transcription using the SuperScript III First-Strand Synthesis SuperMix for qRT-PCR (Invitrogen). Gene expression was quantified using real time qPCR in combination with gene specific primers and the SYBR GREEN system (Roche). The reactions were performed on an Eppendorf Realplex4 cycler (Eppendorf). All samples (n = 3) were run in duplicate. Values were normalized to GAPDH expression. The following primers were used: GADPH-fw, 5′-GAACGGGAAGCTTGTCATCAA-3′; GADPH-rev, 5′-ATCGCCCCACTTGATTTTGG-3′; GFAP-fw, 5′-GAGAA CCGGATCACCATTCC-3′; GFAP-rev, 5′-CCCAGTCTGGAGCA ACCTAC-3′; LCN2-fw, 5-GTTACCTCGTCCGAGTGGTG-3′; LCN2-rev, 5′-TTGGTTCTCCCGTAGAGGGT-3′; VIM-fw, 5′-CTC CGGGAGAAATTGCAGGA-3′; VIM-rev, 5′-TTCAAGGTCAA GACGTGCCA-3′.
