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. Author manuscript; available in PMC: 2012 Jun 1.
Published in final edited form as: Mol Cell Neurosci. 2011 Feb 19;47(2):79–92. doi: 10.1016/j.mcn.2011.02.007

Fig. 3.

Fig. 3

Internalized fluorescent BTX remains associated with 5-HT3A/BBS receptors. a) Surface 5-HT3A/BBS receptors were labeled with BTX/488 and incubated at 37 °C for 30 min. Every 5 min a sample was taken and the cell membranes were labeled with BTX/555 then fixed with 4% formaldehyde. BTX/488 fluorescence was averaged along the WGA/555 fluorescence. b) HEK-293 cells stably expressing 5-HT3A/BBS receptors were transiently transfected with cDNA encoding a 5-HT3A receptor tagged with an HA epitope. Surface receptors were labeled with BTX/488 and then incubated for 30 min at 37 °C. Cells were fixed with 4% formaldehyde for 20 min at 4 °C, permeabilized and incubated with HA antibodies conjugated to Alexa 594 (red label). Arrows indicate co-localized puncta containing both BTX/488 and anti-HA label. Scale bar=10 μm.