Figure 7. ZDHHC14 is required for Kv1-type potassium channel targeting to the AIS in hippocampal neurons.
Hippocampal neurons were transduced on DIV9 with lentiviruses to express GFP without (Control; A) or with Zdhhc14 shRNA#1 (B) and fixed on DIV16 to detect Kv1.1 (left column and green in merged images on right), AnkG (AIS marker, middle column and magenta in merged images on right) and GFP (marker of infected neurons, blue in merged images, right column). Lower panels show magnified view of red or white dashed boxed area in upper panels. (C and D) integrated density and mean gray value, respectively, of Kv1.1 signal within AnkG-defined AIS from images from A, normalized to control condition (C: Mann Whitney test *p=0.020, N = 3 independent cultures, U = 1.00). (D: Mann Whitney test *p=0.020, N = 3 independent cultures, U = 1.00). (E and F) as in A and B, except that antibodies were used to detect Kv1.2, AnkG, and GFP. (G and H) as in C and D but for Kv1.2 AIS targeting from images from E and F (G: Mann Whitney test p=0.029, N = 4 independent cultures, U = 0.00; H: Mann Whitney test p=0.45, N = 4 independent cultures, U = 5.00). (I and J) as in A and B, except that antibodies were used to detect Kv1.4, AnkG, and GFP. (K and L) as in C and D but for Kv1.4 AIS targeting from images from I and J (K: Mann Whitney test *p=0.029, N = 3 independent cultures, U = 0.0; L: Mann Whitney test *p=0.029, N = 3 independent cultures, U = 0.0). Scale bar in full and magnified views: 10 μm. Arrowheads in all panels indicate additional non-magnified AISs in the image. An additional set of example images is provided in Figure 7—figure supplement 1.
Figure 7—figure supplement 1. Additional example images showing that ZDHHC14 is required for Kv1-type potassium channel targeting to the AIS in hippocampal neurons.
