Fig. 3 | Neuropsychopharmacology

Fig. 3

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

Fig. 3

Tubastatin-A slows the lateral mobility of membrane localized GFP-Gαs after photobleaching and enhances isoproterenol elicited cAMP accumulation. C6 cells stably expressing GFP-Gαs  a were treated with the indicated compound and subjected to FRAP analysis as described in the Methods section. Half-time of recovery for GFP-Gαs is increased after treatment (3 days) with tubastatin-A (10 µM), escitalopram (10 µM), imipramine (10 µM), and aripiprazole (500 nM) b, c, and various other antidepressant drugs of different chemical and functional classes [41]. Sample size represents the number of cells assayed, with a minimum of 48 and a maximum of 84 cells assayed per experiment. Data were analyzed by Kruskal–Wallis test (F = 8.767) followed by Dunn’s multiple comparison test. Data are represented as mean ± SEM. (*p < 0.05 compared to vehicle control; **p < 0.01 compared to vehicle control; ****p < .0001 compared to vehicle control). In order to measure cAMP accumulation, C6 cells were grown on glass bottom microscope dishes, pretreated with/without tubastatin-A (3 days) were infected with (1.09 × 109 VG/mL) cADDIS BacMam virus encoding the green “up” cAMP sensor and grown for 24–26 h before live imaging under a ×40 objective on a Zeiss 880. Cells were serum starved with 1% serum for 2–3 h before Isoproterenol (1 μM) treatment. Images were taken every 30 s. Average responses from 4 to 10 cells were selected from the visual field and fluorescence was normalized to baseline fluorescence for each experiment. The montage shown here d is representative of four replicate experiments and the fluorescence values, e, from each experiment were quantified, corrected for baseline fluorescence (∆F/F0), and the data are represented as mean ± SE ****p < 0.0001 e

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