The Journal of Neuroscience, August 24, 2005, ():
5-HT7 Receptor Is Coupled to G
Subunits of Heterotrimeric G12-Protein to Regulate Gene Transcription and Neuronal Morphology
J. Neurosci. Kvachnina et al.
25: 7821
HTML Page - index.htslp
Supplemental Material
Files in this Data Supplement:
- Supplemental Movie
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Video 1. For life-time imaging, NIH3T3 cells transfected with pTracer/5-HT7 vector were starved at 14-16 h post-transfection for 24-36 h and analyzed by laser scanning confocal microscopy. Experiments were performed at 37°C in circulated bicarbonate/CO2 buffered extracellular buffer (150 mM NaCl, 5 mM KCl, 10 mM glucose, 10 mM HEPES, 2 mM CaCl2,1 mM MgCl2, pH 7.4, 330 mOsm). Images were collected with a 63x objective by water immersion under lambda mode setting after recording of corresponding reference spectra. Images were scanned every 5 min during 1 hour. After two scanning cycles the buffer was supplemented with specific agonists (1µM 5-HT). Video-related images are shown in Figure 6B.
- Supplemental Fig. 1
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Figure S1. Distribution of G?12 and G?13 proteins in hippocampal neurons. Endogenous G?12 or G?13 subunits were visualised by immunostaining with specific antibodies. Primary antibodies directed against G?13 were from rabbit, while anti-G?12 from goat. Secondary antibodies donkey anti-goat (AlexaFluor546 from Molecular Probes, diluted 1:1000) and donkey anti-rabbit (Cy2 from Dianova, diluted 1:400) were used for the visualization. Cells were analyzed by laser scanning confocal microscopy (LSM510-Meta) with a 63x objective by water immersion. Scale bar = 10 µM.
