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. 2009 Jun 18;23(9):1466–1478. doi: 10.1210/me.2009-0119

Figure 2.

Figure 2

Phospho-TRH receptor traffics with arrestin. A–C, Cells expressing 2HA-AP-TRH receptor and arrestin-GFP (green) were incubated with mouse anti-HA antibody to label surface receptors and then stimulated with 100 nm TRH for the indicated times. After fixation, cells were stained with antimouse Alexa Fluor 647 antibody to identify anti-HA antibody-labeled receptors (blue) or rabbit anti-phospho-TRH receptor antibody and antirabbit Alexa Fluor 555 antibody to identify phospho-TRH receptors (red). Pink shows overlay of red and blue channels; white shows overlay of all three channels. D and E, Cells expressing 2HA-AP-TRH receptor, arrestin-FLAG, and GFP-Rab5 were stimulated with 100 nm TRH for the indicated times and fixed and stained. Mouse anti-FLAG antibody followed by antimouse Alexa Fluor 647 antibody was used to label arrestin-FLAG (blue). Bar, 10 μm. Arr-, Arrestin; s, seconds; TRHR, TRH receptor.