Skip to main content
. 2022 Jan 5;42(1):2–15. doi: 10.1523/JNEUROSCI.1132-21.2021

Figure 8.

Figure 8.

AnkR binds to Kv3.3 K+ channels and is critical for its proper membrane localization in Purkinje neurons. A, Immunostaining of AnkR (red) and Kv3.3 (green) in sagittal cerebellum of Ank1F/F. Scale bar: 25 µm. B, Immunoblots of urea sample buffer (USB) input, reducing sample buffer (RSB) input, Kv3.3, and AnkR DSP-crosslinked immunoprecipitation reactions using antibodies against AnkR and Kv3.3. C, Immunoblots of AnkR-GFP immunoprecipitations in HEK293T cells coexpressing AnkR-GFP with full-length Flag-tagged Kv3.3a, truncated (ΔEDCPAI) Flag-tagged Kv3.3a, full-length Flag-tagged Kv3.3b, or truncated (ΔEDCPAI) Flag-tagged Kv3.3b. D, Immunostaining of Calbindin (red) and Kv3.3 (green) in sagittal cerebellum of three-month-old Ank1F/F and Ank1F/F;Nestin-Cre. Scale bar: 25 µm. E, Normalized quantification of Kv3.3 expression at the Purkinje neuron soma by measuring the fluorescence intensity of a line across the PCL and dividing by length in three-month sagittal cerebellum of Ank1F/F (N = 3; 1.00 ± 0.04 AU), and Ank1F/F;Nestin-Cre (N = 3; 0.76 ± 0.004 AU). F, Normalized quantification of Kv3.3 expression in the granule cell layer Ank1F/F (N = 3; 1 ± 0.05 AU), and Ank1F/F;Nestin-Cre (N = 3; 0.68 ± 0.05 AU). Error bars indicate mean ± SEM. ML, molecular layer; PCL, Purkinje neuron layer; GCL, granule cell layer.