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. 2023 Oct 19;18(11):2222–2239. doi: 10.1016/j.stemcr.2023.09.011

Figure 3.

Figure 3

Increased miniature excitatory postsynaptic current (mEPSC) amplitudes and elongation of axon initial segment (AIS) following TTX treatment

(A–D) Representative mEPSC traces of vehicle-treated (Veh) and 48-h TTX-treated (TTX) neurons (Ctrl1; A). Quantification of the amplitude (B) and frequency of mEPSCs (C) in Veh- and TTX-treated neurons. n = number of cells/batches: Veh n = 21/3, TTX n = 23/3. Rescaled cumulative mEPSCs amplitude (Scaling = TTX values 1.21; D).

(E) Representative images of vehicle-treated (Veh) and 48 h TTX-treated (TTX) neurons (Ctrl1) stained for MAP2 (gray), surface GluA2 (green), and Synapsin 1/2 (red) (scale bar, 5 μm).

(F) Quantification of GluA2 puncta (number per 10 μm). n = number of cells/batches: Veh n = 18/2, TTX n = 28/2.

(G) Representative images of vehicle-treated (Veh) and 48-h TTX-treated (TTX) neurons (Ctrl1) stained for MAP2 (gray), Homer1 (green), and Synapsin 1/2 (red) (scale bar, 5 μm).

(H) Quantification of Synapsin/Homer1 co-localized puncta (number per 10 μm). n = number of cells/batches: Veh n = 20/1, TTX n = 25/1.

(I) Representative images of vehicle-treated (Veh) and 48 h TTX-treated (TTX) neurons (Ctrl1) stained for MAP2 (gray), ANKG (green), and Hoechst (blue) at DIV 49 (scale bar, 5 μm).

(J and K) Plots showing cumulative fraction and quantification of the length axon initial segment (AIS; μm). n = number of cells/batches: Veh n = 14/2, TTX n = 14/2. Data represent means ± SEM. ns: not significant, p < 0.05, ∗∗∗∗p < 0.0001, unpaired Student’s t test was performed between two groups. All means, SEMs, and test statistics are listed in Table S3.