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. 2005 Jun 7;388(Pt 3):803–811. doi: 10.1042/BJ20050030

Figure 1. Detection of the PDE4D3 splice variant in the rat thyroid cell line FRTL5.

Figure 1

(A) Cytosolic extracts of FRTL5 were immunoprecipitated using a PDE4D3-specific antibody (α-4D3), the antibody M3S1 that recognizes all PDE4D variants (α-4DPAN) and IgG as a negative control. The graph shows the PDE activity of the immunoprecipitation pellets determined at 1 μM cAMP. (B) A cytosolic extract of FRTL5 cells, as well as recombinant rat PDE4D3, were loaded on to SDS/PAGE and subsequently detected by Western blotting using an antibody that recognizes all PDE4 isoforms (α-4PAN), a pan-specific PDE4D antibody (α-4DPAN) and a PDE4D3 splice-variant-selective antibody (α-4D3). Identical amounts of FRTL5 extract and recombinant PDE were loaded for each of the three Western blots. (C) An agarose gel of PDE4D splice-variant-specific DNA fragments amplified from a FRTL5 cDNA using 35 PCR cycles. The identity of the PCR products was confirmed by restriction digestions (results not shown).