FIGURE 2.

Real-time PCR reveals the expression of CaMKIIδ in mouse IHCs. (A–C) Cytoplasm of 3–5 IHCs per sample of P14 mice were collected and analyzed by PCR for the mRNA expression of CaMKIIα, β, γ, and δ. TaqMan assays for bassoon (Bsn) and TATA-binding protein (TBP) were used to control for proper cDNA quality. In three out of three samples, SYBR green fluorescence indicates the expression of CaMKIIδ mRNA; in one sample (C) CaMKIIγ mRNA was expressed in addition. We did not find CAMKIIα or β transcripts in any of the samples. (D) Melting curve analysis (derivative of melting curve is displayed) for the SYBR green assays of the three IHC cDNA samples and brain cDNA samples for comparison; the amplicons using brain cDNA as template revealed the proper melting temperature of the CaMKIIδ and γ amplicons from IHC samples. (E) Amplicons from positive control experiments on brain cDNA, amplicons from experiments in (A–C) and one representative bath control were analyzed by electrophoresis on 2% agarose gels with EtBr staining. Correct sized amplicons were found for bassoon and TBP in the brain and in all three IHC samples, but not in the bath control. Primers for CaMKIIα and CaMKIIβ did not give PCR products of correct size in samples 1–3. CaMKIIγ could be amplified from sample 3 only. The transcripts for CaMKIIδ were present in samples 1, 2, and 3, displayed by amplicons of the correct size.