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. 2014 Mar 25;289(18):12896–12907. doi: 10.1074/jbc.M114.550517

FIGURE 6.

FIGURE 6.

Up-regulated CHOP impairs forward trafficking of GABAB receptors. A, neurons were transfected with either EGFP and HA-tagged GABAB2 (Control), EGFP, HA-GABAB2 and CHOP, or EGFP HA-GABAB2 and CHOPΔLZ (CHOPΔLZ). Twenty-four hours after transfection, GABAB receptors newly inserted into the plasma membrane were visualized by staining with HA-antibodies. Left panel, representative images. Scale bar = 5 μm. Right panel, quantification of GABAB receptor membrane insertion. Data are means ± S.E., 25 neurons/condition from three experiments. ***, p < 0.001; one-way analysis of variance, Dunnett's post-hoc test. B, neurons were treated with thapsigargin (TG) to up-regulate CHOP und subjected to an immunofluorescence-based forward trafficking assay to determine the amounts of GABAB receptors inserted into the plasma membrane within a time period of 16 h (red clusters in representative images, scale bar = 5 μm). CHOP expression is depicted in blue. Cultures not treated with thapsigargin served as controls. Right panel, quantification of GABAB receptor membrane insertion. Data are means ± S.E., 38 (control) and 33 (thapsigargin) neurons from three experiments. ***, p < 0.001; Student's t test.