Figure 7. Reduction of gephyrin phosphorylation at synapses but not within SSDs by 4‐AP.

• A–C
  Reduced immunoreactivity of S270 phosphorylated gephyrin but not total gephyrin levels after 4‐AP treatment, revealed by conventional fluorescence microscopy (KS test, P < 0.001 in B1 and P < 0.0001 in B2, P < 0.0001 in C). Total gephyrin was probed with polyclonal rabbit primary antibody (rbGPHN), and pS270 phosphorylated gephyrin with monoclonal mouse primary antibody (m7a). Number of synapses: n = 4,040 in TTX and n = 3,818 in 4‐AP conditions from two independent experiments. Scale bar: 2 µm.
• D–F
  Reduced numbers of pS270 gephyrin (m7a) detections were recorded by dSTORM for the entire synaptic area (D, KS test, P < 0.01). The number of detections of pS270 gephyrin per SSD (E, KS test, P = 0.18) and the number of SSDs (F, MW test, P = 0.11) were not changed by 4‐AP treatment. Gephyrin was probed with mAb7a antibody and Alexa 647 dye. Number of synapses: n = 810 in TTX and n = 727 in 4‐AP conditions from three independent experiments; (F) mean ± SEM. Scale bar: 100 nm.

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