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. 2009 Nov 18;35(3):792–805. doi: 10.1038/npp.2009.188

Figure 4.

Figure 4

HDAC inhibitors VPA and TSA increase GLT-1 gene transcription in a dose-dependent manner. GLT-1 transcript levels were quantified by real-time PCR after 24 h treatment with 2.5, 5, and 10 mM VPA (a) and with 5, 15, or 30 nM TSA (b) and 24 and 72 h after 1 mM NaB treatment (d). Real-time PCR analysis was carried out with total RNA derived from treated and untreated (Co) cortical astrocytes and with GLT-1- and actin-specific primers. (control is set to 1; bars depict mean±SE (n=3) of the fold-increase; t-test: *p<0.05 vs control, **p<0.005 vs control.) (c) Reporter gene assays were conducted to directly assess promoter activity. Cortical primary astrocytes were transiently transfected with reporter plasmid containing a homolog of the human GLT-1 promoter coupled to a luciferase gene. The cells were cotransfected with nonsecretory Gaussia luciferase expression vector for normalization. The luciferase activity was assayed 24 h after applying drugs at the indicated concentrations. Bars present mean±SE (n=3–5) of the fold-increase compared with untreated cells (set to 1); t-test: *p<0.05 vs control.