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Figure 1 | BMC Cell Biology

Figure 1

From: Epidermal growth factor receptor levels are reduced in mice with targeted disruption of the protein kinase A catalytic subunit

Figure 1

Effect of PKA Cα or Cβ ablation on EGFR expression. (a) Comparison of liver from wild type and PKA Cα and Cβ ablated mice. PKA Cα KO mice showed a clear uniform reduction in size. (b) Confocal immunofluorescence microscopy of frozen liver sections stained by sheep anti-EGFR and Cy3-conjugated donkey anti-sheep antibodies. (c) Western immunoblotting analysis of EGFR expression in liver and brain from wt (+/+), heterozygote (-/+), and Cα and Cβ KO (-/-) mice. Immunoblots were incubated with sheep anti-EGFR, rabbit anti-pan PKA C and mouse anti-PKA C, and rabbit anti-erbB2. Secondary HRP-conjugated anti-IgG antibodies were used for detection. (d) PKA kinase activity in mouse liver. Activity was assayed by phosphorylation of the PKA-specific substrate Kemptide using γ-[32P]ATP. The assay was performed in the presence of cAMP. Activity was measured by liquid scintillation in 3 ml Opti-fluor. Values are given as counts per minute (cpm).

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