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. Author manuscript; available in PMC: 2009 Nov 1.
Published in final edited form as: Nat Med. 2009 May 3;15(5):509–518. doi: 10.1038/nm.1962

Figure 4. Detection and quantification of Isoform 3.1 mRNA and protein.

Figure 4

(A) PCR of KCNH2-1A and Isoform 3.1 using isoform specific primer pairs in human heart, hippocampus, and fetal brain tissue extracts. PCR products for KCNH2-1A correspond to 373 and 463 bps, respectively, and for Isoform 3.1 at 388 and 478 bp, respectively. The band ∼1.5 kb in Iso1-Ex4 lanes corresponds to small amounts of unspliced RNA or genomic DNA contaminant present in the samples. (B) Protein expression of KCH2-1A and Isoform 3.1 in human, primate, and mouse frontal cortex. Positive control lanes include HEK cells transfected with full-length KCH2-1A (WT) and Isoform 3.1. Proteins extracted from human brain regions show two distinct bands, one equivalent in size to isoform 3.1 transfected HEK cells. However, the larger protein band observed in human brain did not correspond to the size of KCNH2-1A transfected HEK proteins. Instead, the larger band occurs at ∼160 kDa, which is the reported size of KCNH2-1A from in vivo protein extracts suggesting post-translational modifications. (D) Primary rat neurons transfected with KCNH2-1A or Isoform 3.1 containing vectors.