Figure 5: Late-stage AP-2 depletion does not impair synaptic function.

(a,b) Representative images of axon extension in hippocampal neurons (cultured for 14–16 days) expressing GFP-Tau with as an axonal marker taken from: (a) Control cell (expressing GFP-Tau alone), (b) Late AP-2KD cell (expressing GFP-Tau + shRNA targeting μ2 subunit of AP-2). Knockdown shRNA was added at 8 DIV after plating. A-B (Inset), magnified cell body of neurons that were immunolabeled with left to right: anti-GFP (green) and anti-α-adaptin antibody (red) and merged (overlay of red and green imaged). (c) Representative 1 action potential stimulated GCaMP6f traces in control (black) and late AP-2KD neurons (red). (d) Mean value of Physin-GCaMP6f 1 action potential response amplitudes in control and AP-2KD neuron. Con = 2.24 ± 0.64% (n = 4 cells), AP-2KD = 2.02 ± 0.24% (n = 4 cells). (e) Representative vG-pH traces of control and late AP-2KD neurons 100 action potential pulses at 10 Hz. (f) Mean values of peak vG-pH in control and AP-2KD cells normalized to NH₄Cl response. Con = 27.98 ± 0.01% (n = 11 cells), AP-2KD = 24.98 ± 4.58% (n = 8 cells), p = 0.51.