Figure 2.

Regulation of mRNA and protein expression of cfos and fosB or protein content of Caveolin-1 (Cav-1) by acute treatment with fluoxetine in astrocytes. (A,B) After pretreatment in serum-free medium with or without U0126, an inhibitor of MEK or LY294002, an inhibitor of PI3K for 15 min, cells were incubated for 1 or 4 h in the absence of any drug (0 μM fluoxetine, control) or in the presence of 0.1, 0.5, 1, 5, or 10 μM fluoxetine. (A) Southern blot from representative experiment. Similar results were obtained from three independent experiments. Average mRNA expression was quantitated as ratios between c-fos and TBP, used as a house-keeping gene and between fosB and TBP. (B) Immunoblot from a representative experiment. Similar results were obtained from three independent experiments. Average protein expression was quantitated as ratios between c-Fos and β-actin, used as a house-keeping gene and between FosB and β-actin. SEM values are indicated by vertical bars. *Indicates statistically significant (P < 0.05) difference from 0, 0.1, 0.5 and 1 μM fluoxetine; **indicates statistically significant (P < 0.05) difference from all other groups but not from each other. (C) Cells were incubated for 1 or 4 h in the absence of any drug (0 μM fluoxetine, control) or in the presence of 0.1, 0.5, 1, 5, or 10 μM fluoxetine. Left panel shows the immunoblot from a representative experiment. Similar results were obtained from three independent experiments. Average protein expression was quantified as ratio between Cav-1 and β-actin, used as a house-keeping gene (right panel). SEM values are indicated by vertical bars.