Pharmacological inhibition of CK2 releases ion channels from immobility and destabilizes ion channel concentration at the AIS in young neurons. (A–D) SPT on Kv-Nav (A and B) and Kv-Nav 4SA (C and D) in DIV 4 neurons in the presence of the CK2 inhibitor DMAT. QD trajectories were recorded at the AIS of DIV 4 neurons acutely treated with 50 µM DMAT or a vehicle only during the indicated time intervals. For Kv-Nav, trajectories for vehicle (n = 14) and DMAT (n = 19) were analyzed from three and four independent experiments, respectively. For Kv-Nav 4SA, trajectories for vehicle (n = 11) and DMAT (n = 12) were analyzed from three indepent experiments. MDC (25–75% IQR) before and after drug addition for Kv-Nav (A and B) and Kv-Nav 4SA (C and D). WSR: ***, P < 0.001. (E) Effect of DMAT on the accumulation of AIS components in young neurons. DIV 3 cells were treated with either 50 µM DMAT or with DMSO (control cells) for 4, 8, and 24 h and were subsequently immunostained for Nav1 channels and ankG. Quantification of the respective fluorescence intensity was achieved and was normalized to 100%, representing the staining intensity measured in control cells. Histogram represents the normalized mean values ± SEM. The number of quantified AISs range from 69 to 118 per condition (two independent experiments). t test: *, P < 0.05; **, P < 0.01; and ***, P < 0.001.