Fig. 1 | Neuropsychopharmacology

Fig. 1

From: Disruption of lipid-raft localized Gαs/tubulin complexes by antidepressants: a unique feature of HDAC6 inhibitors, SSRI and tricyclic compounds

Fig. 1

Sustained treatment with tubastatin-A, escitalopram, or imipramine induce translocation of Gαs out of lipid-raft domains, but only tubastatin-A increases acetylation of α-tubulin. C6 glioma cells were treated (3 days) with either tubastatin-A (10 µM), escitalopram (10 µM), imipramine (10 µM), or vehicle control and cells were collected for isolation of lipid-raft domains using sucrose density gradient centrifugation. Ten different fractions were collected (A&E). Lipid-raft domains (fractions 3–5) were revealed via enrichment of Caveolin-1, a lipid-raft protein. The collected membrane fractions (1–10), cell homogenate (H) and isolated plasma membrane (PM) were prepared for SDS-PAGE and membranes were incubated with Cav1, acetylated-α-tubulin, total α-tubulin and Gαs antibodies. Three replicate experiments were conducted and Gαs, acetylated-α-tubulin and total tubulin from Homogenate (H), Plasma Membrane (PM), and lipid-rafts were quantified and plotted as percent of Gαs and fold change in acetylation compared to control samples b, c, d, f, g, h. Data in b, c, d were analyzed using unpaired t-test followed by Welch’s correction and mean ± SEM are represented as fold change or percent change compared to controls. Each condition was compared to vehicle control by unpaired t-test. The p-values obtained from this test were used for the represented graphs in each figure. Data in figure f, g, h were analyzed by one way ANOVA for comparisons between Control, Escitalopram, and Imipramine followed by Kruskal–Wallis test (*p < 0.05 compared to vehicle control; **p < 0.01 compared to vehicle control, ***p < 0.001 compared to vehicle control)

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