Fig. 5. ß₂ARs localize with EGFP-rab11WT and pulse-chased transferrin following a prolonged exposure to agonist. (A) 12ß6 cells transiently expressing empty EGFP vector (a,a') or EGFP-rab11WT (b,b') were treated with ISO for 6 hours, fixed immediately, and ß₂ARs and EEA1 labeled as described. ß₂ARs appear red and EEA1 appear green. The arrows in panel b indicates ß₂ARs that localize with EGFP-rab11WT. (B) 12ß6 cells transiently expressing empty EGFP vector were treated with ISO for 6 hours. During the last 30 minutes of ISO treatment prior to fixation, cells were fed Alexa 594-transferrin for 15 minutes, washed, and the transferrin chased for 15 minutes in the continuous presence of ISO. ß₂ARs are shown in green and Alexa 594-transferrin appears red. Areas of colocalization of receptors and transferrin appear yellow, as shown by the arrow. Scale bar, 10 µm.