Figure 4.

Inhibition of Tpm3.1 Reduces the Accumulation of Ankyrin G at the AIS

(A) Rat hippocampal neurons treated at 10 DIV using DMSO or the small-molecule Tpm3.1 inhibitors TR100 or Anisina (ATM3507) for 2, 3, or 6 h. Anti-MAP2 served to label the somatodendritic domain; anti-ankyrin G served to measure the accumulation of ankyrin G. Arrows point to axons.

(B) Smoothed ankyrin G fluorescence intensity line profiles (gray lines) along each neurite of the corresponding cell in (A), normalized to the median peak value (black line).

(C) AIS localization indices for neurons treated using DMSO, TR100 (10 or 15 μM), or Anisina (5 or 7.5 μM) for 2, 3, or 6 h. All treatment groups were significantly different from DMSO controls (Mann-Whitney U test; DMSO 0.2%, 2 h: 0.94 ± 0.006, mean ± SEM; DMSO 0.2%, 3 h: 0.93 ± 0.006; DMSO 0.2%, 6 h: 0.96 ± 0.005; TR100 10 μM, 2 h: 0.59 ± 0.050, p < 0.001; TR100 10 μM, 3 h: 0.50 ± 0.051, p < 0.001; TR100 10 μM, 6 h: 0.42 ± 0.066, p < 0.001; TR100 15 μM, 2 h: 0.45 ± 0.060, p < 0.001; TR100 15 μM, 3 h: 0.42 ± 0.060, p < 0.001; TR100 15 μM, 6 h: 0.35 ± 0.056, p < 0.001; Anisina 5 μM, 2 h: 0.58 ± 0.055, p < 0.001; Anisina 5 μM, 3 h: 0.49 ± 0.049, p < 0.001; Anisina 5 μM, 6 h: 0.44 ± 0.039, p < 0.001; Anisina 7.5 μM, 2 h: 0.51 ± 0.041, p < 0.001; Anisina 7.5 μM, 3 h: 0.41 ± 0.062, p < 0.001; Anisina 7.5 μM, 6 h: 0.36 ± 0.075, p < 0.001; for each treatment, n = 12, 3 independent experiments). The mean ALI of the treatment groups was negatively correlated with treatment duration and concentration. Black circles represent mean values. Box borders represent the 25^th and 75^th percentiles, whiskers represent minimum and maximum values less than 1.5x the interquartile range lower or higher than the 25^th or 75^th percentiles, respectively (Tukey style). Dotted lines connect mean values. Scale bar: 5 μm.

See also Figure S5.