Fig. 6: Acute degradation of GephyrinA29:mAID:HT is followed by loss of GABARs at the same synapses. | Communications Biology

Fig. 6: Acute degradation of GephyrinA29:mAID:HT is followed by loss of GABARs at the same synapses.

From: Revealing acute consequences of rapid degradation of synaptic fusion proteins at individual synapses using Auxin-Inducible Degron 2 technology

Fig. 6: Acute degradation of GephyrinA29:mAID:HT is followed by loss of GABARs at the same synapses.

A Top panels: A rat cortical neuron in culture co-expressing GephyrinA29:mAID:HT (labeled with JF635-HT), SEpH:GABAARα2 and OsTIR1-P2A-mCherry. Bottom panels: Region in yellow rectangle at greater detail, before, and after addition of 5-Ph-IAA. Note that the near complete loss of JF635-HT fluorescence (presumably reflecting GephyrinA29:mAID:HT degradation) is associated with a detectable reduction in SEpH:GABAARα2 fluorescence. Scale bars: 10 µm (top panels) 5 µm (bottom panels). B JF635-HT fluorescence measured at 50 synapses of each neuron tracked throughout the experiments. Each thin gray line is the average JF635-HT fluorescence measured for the synapses of one neuron (900 synapses from 18 neurons from 3 experiments). Thick magenta line is the population average. C changes in SEpH:GABAARα2 fluorescence at the same synapses and neurons of (B). Thick brown line is the population average. D SEpH:GABAARα2 fluorescence measured at 150 synapses of neurons positive for SEpH:GABAARα2 and OsTIR1-P2A-mCherry but negative for GephyrinA29:mAID:HT. Each thin gray line is the average fluorescence measured for the synapses of one neuron (3 neurons from 2 experiments). Thick gray line is the population average. E Pooled data. Error bars are standard deviations. Test for difference between GephyrinA29:mAID:HT positive and negative cells – unpaired t-test, without assuming equal variances; applied to data of last time point.

Back to article page