Fig. 6 | Scientific Reports

Fig. 6

From: The E3 ubiquitin ligase MARCH1 mediates downregulation of plasma membrane GABAB receptors under ischemic conditions by inhibiting fast receptor recycling

Fig. 6

M1-Pep restored glutamate induced loss of cell surface GABAB receptors. Neurons were transfected with GABAB2 tagged at the N-terminus with the minimum binding site for α-bungarotoxin (GABAB2(BBS)) together with EGFP (for quick identification of transfected neurons) and GABAB1. Cell surface receptors containing GABAB2(BBS) were labeled with AlexaFluor 555 conjugated α-bungarotoxin and then neurons were treated with 50 µM glutamate to induce downregulation of the receptors. Neurons were analyzed by live cell imaging for internalization of labeled receptors. M1-Pep and Crtl-Pep were added 13 min after adding glutamate. Top: Representative images (scale bar: 10 μm). Bottom: Quantification of the immunofluorescence signal and statistical analysis. The statistical evaluation was performed on data recorded 15 min after glutamate treatment. The data represent the mean ± SD of 4–6 neurons derived from three independent experiments. One-way ANOVA followed by Tukey’s multiple comparison test (ns, p > 0.05; **p < 0.01).

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