The Journal of Neuroscience, September 8, 2004, ():
KChIPs and Kv4 
Subunits as Integral Components of A-Type Potassium Channels in Mammalian Brain
J. Neurosci. Rhodes et al.
24: 7903
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplemental Fig. 1: Immunoblots demonstrating the specificity of the anti-Kv4 and anti-KChIP mAbs. Detergent lysates of COS-1 cells transiently transfected with Kv4.2, Kv4.3 or the indicated KChIP cDNAs were fractionated by SDS-PAGE and transferred to nitrocellulose, as described (Methods). The resulting immunoblots were probed with the indicated antibody, together with a mouse mAb against ? actin. The presence and intensity of the actin-immunoreactive bands indicates that roughly equal amounts of cell lysates were loaded in each lane. In the immunoblots for Kv4.2 and Kv4.3, the antibodies detect a major band at approximately 70 kDa, respectively, plus higher molecular weight bands corresponding to aggregated Kv4 ? subunits (asterisks, see also Sheng et al., 1993; An et al., 2000). Importantly, the K57/27 anti-Kv4.2 mAb does not recognize Kv4.3, and the K75/30 anti-Kv4.3 antibody does not recognize Kv4.2. Similarly, each of the KChIP-specific antibodies recognizes only the cognate antigen. In lysates prepared from KChIP3 transfected cells, we see a higher, ~30kDa, and a lower, ~25kDa band.
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Supplemental Fig. 2. Gradient of Kv4.2 localization within hippocampal dentate granule and CA1 pyramidal dendrites. Confocal images showing distribution of Kv4.2 and PSD-95 in the dentate gyrus (left panels) and CA1 (right panels). Note dramatic gradient of Kv4.2 staining intensity relative to that of PSD-95 in the molecular layer of dentate gyrus, consistent with the immunoperoxidase staining shown in Fig. 3. A more subtle gradient of Kv4.2 staining relative to that of PSD-95 is also seen in stratum radiatum of CA1. Abbreviations: ig = infragranular layer; gc = granule cell layer; ml = molecular layer, or = stratum oriens; py = stratum pyramidale; rd = stratum radiatum; ml = stratum lacunosum-moleculare.
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Supplemental Fig. 3. Colocalization of KChIP1 and calretinin in hippocampal interneurons. Confocal images show immunofluorescence labeling for KChIP1 (panel A, red in panel C) and calretinin (panel B, green in panel C) in stratum pyramidale of CA1. Note extensive cellular overlap of membrane-associated KChIP1 and cytoplasmic calretinin staining.
