Fig. 2

Validation of selected upregulated genes. a Transcriptional levels of MAP2K5, GRP65, and CXCL5 in astrocytes stimulated with 1, 10, and 100 μM of Meth for 24 h, examined by SyBrGreen qRT-PCR. Results are the Mean ± SEM of three independent experiments performed in duplicate. b Representative western blots for detection of MEK5, the protein encoded by MAP2K5 gene, GPR65, and normalizing b-actin, in protein extracts from astrocytes stimulated with 1, 10, and 100 μM of Meth for 24 h. c Normalized band intensity was calculated in ImageJ software (NIH). d Confocal imaging showing representative astrocytic cultures stained with specific antibodies for detection of MEK5, GPR65, and CXCL5 in 10 and 100 μM Meth treatments. e Fluorescence intensity of the expression of MEK5, GPR65, and CXCL5 was calculated in ImageJ (NIH) for cultures stimulated with 1, 10, and 100 μM, as well as controls. *p < 0.05 in one-way ANOVA followed by Bonferroni’s post hoc comparison against control conditions