a | | Gephyrin distance to synapsin1 | Gephyrin positive synapses (UMAP 6 vs UMAP1-5) | Normal | Permutation *t* test | 5000 | 0.002 | −0.44 | −0.46 | −0.40 | −16.60 | −20.65 | −12.49 | 1.0 | * | 6 | 5 |

| 3*A*,*F* | Gephyrin integrated intensity | Gephyrin positive synapses (UMAP 6 vs UMAP1-5) | Normal | Permutation *t* test | 5000 | 0.002 | 0.35 | 0.20 | 0.49 | 2.70 | 1.25 | 5.46 | 1.0 | * | 6 | 5 |

| | PSD-95 distance to synapsin1 | PSD-95 positive synapses (UMAP6 vs UMAP1-5) | Normal | Permutation *t* test | 5000 | 0.003 | −0.56 | −0.70 | −0.47 | −5.67 | −7.51 | −4.11 | 1.0 | * | 6 | 5 |

| 3*A*,*F* | PSD-95 integrated intensity | PSD-95 positive synapses (UMAP6 vs UMAP1-5) | Normal | Permutation *t* test | 5000 | 0.007 | −0.23 | −0.31 | −0.10 | −2.56 | −6.14 | −0.69 | 1.0 | * | 6 | 5 |

| 3*A*,*F* | Homer1-b/c integrated intensity | Homer1-b/c positive synapses (UMAP6 vs UMAP1) | Normal | Permutation *t* test | 5000 | 0.003 | −0.56 | −0.67 | −0.43 | −5.08 | −7.32 | −3.58 | 1.0 | * | 6 | 5 |

b | 4*A* | Excitatory synapse number | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.626 | 2.99 | −7.40 | 15.23 | 0.29 | −1.10 | 1.59 | 0.1 | | 6 | 6 |

| | Inhibitory synapse number | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.592 | 3.62 | −7.56 | 16.24 | 0.33 | −1.03 | 1.69 | 0.1 | | 6 | 6 |

| 4*B* | Excitatory:inhibitory synapse ratio | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.592 | −0.86 | −4.03 | 1.78 | −0.32 | −1.64 | 1.13 | 0.1 | | 6 | 6 |

c | 4*C* | vGlut1 integrated intensity | Untreated vs TTX (excitatory synapses) | Normal | Permutation *t* test | 5000 | 0.006 | 0.17 | 0.10 | 0.24 | 2.82 | 1.17 | 5.03 | 1.0 | * | 6 | 5 |

| | vGAT integrated intensity | Untreated vs TTX (inhibitory synapses) | Normal | Permutation *t* test | 5000 | 0.001 | 0.60 | 0.44 | 0.75 | 4.31 | 2.84 | 6.50 | 1.0 | * | 6 | 5 |

| | Synapsin1 integrated intensity | Untreated vs TTX (excitatory synapses) | Normal | Permutation *t* test | 5000 | 0.110 | 0.10 | −0.02 | 0.18 | 1.11 | −0.52 | 4.00 | 0.4 | | 6 | 5 |

| | Synapsin1 integrated intensity | Untreated vs TTX (inhibitory synapses) | Normal | Permutation *t* test | 5000 | 0.004 | 0.35 | 0.23 | 0.47 | 3.11 | 1.53 | 4.95 | 1.0 | * | 6 | 5 |

| | Bassoon integrated intensity | Untreated vs TTX (excitatory synapses) | Normal | Permutation *t* test | 5000 | 0.129 | 0.18 | −0.04 | 0.38 | 1.02 | −0.66 | 2.49 | 0.3 | | 6 | 5 |

| | Bassoon integrated intensity | Untreated vs TTX (inhibitory synapses) | Normal | Permutation *t* test | 5000 | 0.360 | 0.11 | −0.12 | 0.32 | 0.57 | −0.98 | 2.07 | 0.1 | | 6 | 5 |

| | SHANK3 integrated intensity | Untreated vs TTX (excitatory synapses) | Normal | Permutation *t* test | 5000 | 0.008 | 0.35 | 0.12 | 0.46 | 2.45 | 0.60 | 7.29 | 0.9 | * | 6 | 5 |

| | Homer1-b/c integrated intensity | Untreated vs TTX (excitatory synapses) | Normal | Permutation *t* test | 5000 | 0.037 | 0.15 | 0.02 | 0.26 | 1.54 | −0.02 | 3.44 | 0.6 | * | 6 | 5 |

| | PSD-95 integrated intensity | Untreated vs TTX (excitatory synapses) | Normal | Permutation *t* test | 5000 | 0.017 | 0.15 | 0.06 | 0.24 | 1.93 | 0.71 | 3.31 | 0.8 | * | 6 | 5 |

| | Gephyrin integrated intensity | Untreated vs TTX (inhibitory synapses) | Normal | Permutation *t* test | 5000 | 0.610 | 0.07 | −0.14 | 0.35 | 0.31 | −1.24 | 1.72 | 0.1 | | 6 | 5 |

| | NR2B integrated intensity | Untreated vs TTX (excitatory synapses) | Normal | Permutation *t* test | 5000 | 0.089 | 0.12 | −0.02 | 0.22 | 1.18 | −0.48 | 3.26 | 0.4 | | 6 | 5 |

| | Cortactin integrated intensity | Untreated vs TTX (excitatory synapses) | Normal | Permutation *t* test | 5000 | 0.549 | −0.09 | −0.33 | 0.18 | −0.39 | −2.45 | 1.07 | 0.1 | | 6 | 5 |

| | Cortactin integrated intensity | Untreated vs TTX (inhibitory synapses) | Normal | Permutation *t* test | 5000 | 0.589 | −0.09 | −0.41 | 0.24 | −0.31 | −1.87 | 1.14 | 0.1 | | 6 | 5 |

| | Actin integrated intensity | Untreated vs TTX (excitatory synapses) | Normal | Permutation *t* test | 5000 | 0.475 | −0.07 | −0.26 | 0.04 | −0.53 | −1.80 | 1.06 | 0.1 | | 6 | 5 |

| | Actin integrated intensity | Untreated vs TTX (inhibitory synapses) | Normal | Permutation *t* test | 5000 | 0.794 | 0.04 | −0.22 | 0.38 | 0.17 | −1.48 | 2.06 | 0.1 | | 6 | 5 |

d | 5*C* | Synapsin1 UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.202 | 0.13 | −0.05 | 0.30 | 0.84 | −0.72 | 2.49 | 0.2 | | 5 | |

| | Synapsin1 UMAP2 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.479 | −0.03 | −0.12 | 0.03 | −0.47 | −1.83 | 1.09 | 0.1 | | 6 | 5 |

| | Synapsin1 UMAP3 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.569 | 0.03 | −0.10 | 0.09 | 0.36 | −1.23 | 3.70 | 0.1 | | 6 | 5 |

| | Synapsin1 UMAP4 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.237 | −0.06 | −0.17 | 0.03 | −0.82 | −2.54 | 1.03 | 0.2 | | 6 | 5 |

| | Synapsin1 UMAP5 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.099 | −0.09 | −0.18 | 0.01 | −1.09 | −2.63 | 0.48 | 0.4 | | 6 | 5 |

| | Synapsin1 UMAP6 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.005 | 0.37 | 0.24 | 0.49 | 3.09 | 1.64 | 4.75 | 1.0 | * | 6 | 5 |

| | Gephyrin UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.464 | 0.02 | −0.03 | 0.07 | 0.47 | −0.92 | 1.96 | 0.1 | | 6 | 5 |

| | Gephyrin UMAP2 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.261 | −0.03 | −0.08 | 0.02 | −0.68 | −2.13 | 0.68 | 0.2 | | 6 | 5 |

| | Gephyrin UMAP3 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.345 | −0.03 | −0.08 | 0.02 | −0.61 | −1.96 | 0.76 | 0.1 | | 6 | 5 |

| | Gephyrin UMAP4 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.304 | 0.02 | −0.02 | 0.06 | 0.69 | −1.58 | 2.43 | 0.2 | | 6 | 5 |

| | Gephyrin UMAP5 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.647 | 0.02 | −0.05 | 0.08 | 0.30 | −1.82 | 2.69 | 0.1 | | 6 | 5 |

| | Gephyrin UMAP6 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.733 | 0.03 | −0.12 | 0.25 | 0.20 | −1.41 | 1.56 | 0.1 | | 6 | 5 |

| | Homer1-b/c UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.221 | 0.09 | −0.05 | 0.23 | 0.81 | −0.89 | 2.60 | 0.2 | | 6 | 5 |

| | Homer1-b/c UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.452 | 0.00 | 0.00 | 0.00 | −0.81 | −1.64 | −0.40 | 0.2 | | 6 | 5 |

| | Homer1-b/c UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.438 | 0.05 | −0.07 | 0.16 | 0.49 | −1.17 | 2.16 | 0.1 | | 6 | 5 |

| | Homer1-b/c UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.660 | 0.01 | −0.04 | 0.08 | 0.29 | −1.70 | 2.07 | 0.1 | | 6 | 5 |

| | Homer1-b/c UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.474 | 0.02 | −0.05 | 0.08 | 0.41 | −1.23 | 2.01 | 0.1 | | 6 | 5 |

| | Homer1-b/c UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.031 | 0.19 | 0.07 | 0.29 | 1.80 | 0.48 | 4.20 | 0.8 | * | 6 | 5 |

| | NR2B UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.144 | 0.10 | −0.03 | 0.22 | 0.97 | −0.48 | 2.77 | 0.3 | | 6 | 5 |

| | NR2B UMAP2 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.827 | −0.02 | −0.14 | 0.11 | −0.15 | −1.71 | 1.68 | 0.1 | | 6 | 5 |

| | NR2B UMAP3 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.312 | 0.07 | −0.04 | 0.17 | 0.63 | −0.95 | 2.41 | 0.2 | | 6 | 5 |

| | NR2B UMAP4 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.661 | 0.05 | −0.18 | 0.28 | 0.26 | −1.50 | 2.35 | 0.1 | | 6 | 5 |

| | NR2B UMAP5 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.744 | −0.03 | −0.20 | 0.12 | −0.20 | −1.80 | 1.34 | 0.1 | | 6 | 5 |

| | NR2B UMAP6 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.533 | 0.14 | −0.33 | 0.54 | 0.37 | −1.08 | 1.97 | 0.1 | | 6 | 5 |

| | PSD-95 UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.313 | 0.06 | −0.04 | 0.17 | 0.65 | −0.85 | 2.04 | 0.2 | | 6 | 5 |

| | PSD-95 UMAP2 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.053 | 0.06 | 0.01 | 0.11 | 1.43 | −0.33 | 3.18 | 0.6 | | 6 | 5 |

| | PSD-95 UMAP3 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.778 | 0.02 | −0.10 | 0.15 | 0.17 | −1.28 | 1.56 | 0.1 | | 6 | 5 |

| | PSD-95 UMAP4 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.882 | 0.00 | 0.00 | 0.00 | −0.12 | −1.47 | 1.58 | 0.1 | | 6 | 5 |

| | PSD-95 UMAP5 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.184 | 0.00 | 0.00 | 0.01 | 0.86 | 0.40 | 1.50 | 0.2 | | 6 | 5 |

| | PSD-95 UMAP6 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.729 | −0.01 | −0.08 | 0.07 | −0.21 | −1.69 | 1.26 | 0.1 | | 6 | 5 |

| | SHANK3 UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.018 | 0.23 | 0.11 | 0.35 | 2.07 | 0.81 | 4.11 | 0.9 | * | 6 | 5 |

| | SHANK3 UMAP2 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.011 | 0.10 | 0.04 | 0.15 | 1.94 | 0.55 | 3.24 | 0.8 | * | 6 | 5 |

| | SHANK3 UMAP3 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.019 | 0.20 | 0.08 | 0.30 | 1.85 | 0.55 | 4.35 | 0.8 | * | 6 | 5 |

| | SHANK3 UMAP4 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.186 | 0.06 | −0.03 | 0.14 | 0.85 | −0.62 | 2.44 | 0.2 | | 6 | 5 |

| | SHANK3 UMAP5 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.285 | 0.06 | −0.04 | 0.16 | 0.67 | −0.80 | 2.17 | 0.2 | | 6 | 5 |

| | SHANK3 UMAP6 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.360 | 0.07 | −0.06 | 0.22 | 0.58 | −0.84 | 2.15 | 0.1 | | 6 | 5 |

| | Actin UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.345 | −0.10 | −0.28 | 0.06 | −0.60 | −1.95 | 1.11 | 0.1 | | 6 | 5 |

| | Actin UMAP2 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.927 | 0.00 | −0.05 | 0.06 | −0.06 | −1.54 | 1.59 | 0.1 | | 6 | 5 |

| | Actin UMAP3 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.407 | −0.06 | −0.18 | 0.06 | −0.56 | −2.01 | 1.13 | 0.1 | | 6 | 5 |

| | Actin UMAP4 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.721 | −0.01 | −0.07 | 0.05 | −0.21 | −1.70 | 1.26 | 0.1 | | 6 | 5 |

| | Actin UMAP5 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.923 | −0.01 | −0.18 | 0.07 | −0.11 | −1.72 | 2.14 | 0.1 | | 6 | 5 |

| | Actin UMAP6 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.876 | 0.01 | −0.14 | 0.19 | 0.10 | −1.31 | 1.65 | 0.1 | | 6 | 5 |

| | Bassoon UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.250 | 0.08 | −0.04 | 0.21 | 0.74 | −0.90 | 1.94 | 0.2 | | 6 | 5 |

| | Bassoon UMAP2 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.593 | −0.02 | −0.11 | 0.06 | −0.33 | −1.97 | 1.56 | 0.1 | | 6 | 5 |

| | Bassoon UMAP3 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.064 | 0.09 | 0.02 | 0.22 | 1.16 | −0.33 | 2.12 | 0.4 | | 6 | 5 |

| | Bassoon UMAP4 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.877 | 0.01 | −0.09 | 0.13 | 0.09 | −2.47 | 3.45 | 0.1 | | 6 | 5 |

| | Bassoon UMAP5 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.569 | 0.02 | −0.05 | 0.08 | 0.33 | −1.26 | 2.33 | 0.1 | | 6 | 5 |

| | Bassoon UMAP6 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.281 | 0.07 | −0.06 | 0.19 | 0.64 | −0.83 | 2.20 | 0.2 | | 6 | 5 |

| | Cortactin UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.201 | −0.09 | −0.24 | 0.01 | −0.84 | −2.03 | 0.63 | 0.2 | | 6 | 5 |

| | Cortactin UMAP2 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.573 | −0.02 | −0.11 | 0.05 | −0.34 | −1.81 | 1.07 | 0.1 | | 6 | 5 |

| | Cortactin UMAP3 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.390 | 0.00 | 0.00 | 0.00 | 0.55 | −0.98 | 2.37 | 0.1 | | 6 | 5 |

| | Cortactin UMAP4 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.867 | 0.00 | 0.00 | 0.00 | 0.11 | −1.21 | 1.57 | 0.1 | | 6 | 5 |

| | Cortactin UMAP5 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.781 | −0.02 | −0.12 | 0.11 | −0.16 | −2.09 | 1.78 | 0.1 | | 6 | 5 |

| | Cortactin UMAP6 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.585 | −0.06 | −0.24 | 0.14 | −0.33 | −1.87 | 1.10 | 0.1 | | 6 | 5 |

| | vGlut1 UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.044 | 0.19 | 0.01 | 0.30 | 1.49 | −0.25 | 3.75 | 0.6 | * | 6 | 5 |

| | vGlut1 UMAP2 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.252 | 0.03 | −0.02 | 0.08 | 0.78 | −0.83 | 2.69 | 0.2 | | 6 | 5 |

| | vGlut1 UMAP3 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.045 | 0.08 | 0.01 | 0.16 | 1.38 | −0.39 | 3.00 | 0.5 | * | 6 | 5 |

| | vGlut1 UMAP4 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.231 | 0.03 | −0.02 | 0.07 | 0.77 | −0.83 | 2.44 | 0.2 | | 6 | 5 |

| | vGlut1 UMAP5 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.722 | 0.01 | −0.05 | 0.09 | 0.21 | −2.28 | 3.33 | 0.1 | | 6 | 5 |

| | vGlut1 UMAP6 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.358 | −0.01 | −0.02 | 0.01 | −0.57 | −2.24 | 1.05 | 0.1 | | 6 | 5 |

| | vGAT UMAP1 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.386 | 0.00 | −0.01 | 0.00 | −0.60 | −1.69 | 1.01 | 0.1 | | 6 | 5 |

| | vGAT UMAP2 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.883 | 0.00 | 0.00 | 0.00 | 0.08 | −1.41 | 1.55 | 0.1 | | 6 | 5 |

| | vGAT UMAP3 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.101 | −0.01 | −0.02 | 0.00 | −1.03 | −1.98 | 0.55 | 0.3 | | 6 | 5 |

| | vGAT UMAP4 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.861 | 0.00 | 0.00 | 0.01 | 0.06 | −1.31 | 1.57 | 0.1 | | 6 | 5 |

| | vGAT UMAP5 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.101 | −0.01 | −0.01 | 0.00 | −1.13 | −2.16 | −0.49 | 0.4 | | 6 | 5 |

| | vGAT UMAP6 | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.002 | 0.86 | 0.61 | 1.08 | 3.99 | 2.55 | 6.30 | 1.0 | * | 6 | 5 |

e | 5*C* | UMAP1 synapse number | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.117 | 0.11 | 0.01 | 0.22 | 1.09 | −0.56 | 2.56 | 0.4 | | 6 | 5 |

| | UMAP2 synapse number | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.042 | −0.27 | −0.47 | −0.04 | −1.52 | −3.59 | 0.15 | 0.6 | * | 6 | 5 |

| | UMAP3 synapse number | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.092 | 0.23 | 0.07 | 0.53 | 1.17 | 0.10 | 2.19 | 0.4 | | 6 | 5 |

| | UMAP4 synapse number | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.091 | −0.14 | −0.28 | 0.03 | −1.13 | −3.51 | 0.56 | 0.4 | | 6 | 5 |

| | UMAP5 synapse number | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.002 | −0.42 | −0.60 | −0.30 | −3.12 | −4.32 | −1.96 | 1.0 | * | 6 | 5 |

| | UMAP6 synapse number | Untreated vs TTX | Normal | Permutation *t* test | 5000 | 0.428 | 0.12 | −0.11 | 0.42 | 0.51 | −0.84 | 1.85 | 0.1 | | 6 | 5 |