Fig. 4.

Brain Ca²⁺-dependent PLA₂ enzymatic activity, protein and mRNA expression in wild type and COX-1^−/− mice. (A) Brain Ca²⁺-dependent cPLA₂ and sPLA₂ activity was assessed using cPLA₂ and sPLA₂ activity assay kits. The results are expressed as mean ± SEM of twelve independent samples, each assayed in duplicate. (B) Brain cPLA₂ and sPLA₂ mRNA expression measured by real-time PCR and normalized to an endogenous reference (pgk1) in the COX-1^−/− mice relative to the wild type, using the ΔΔC_T method. The results are means ± SEM of six independent samples, each assayed in duplicate. (C) Representative immunoblots illustrating cPLA₂ (110 kDa), sPLA₂ (14 kDa), and β-actin (42 kDa) proteins. Relative integrated density values of cPLA₂ and sPLA₂ in brain from COX-1^−/− mice are reported as percentage of values in wild type mice. Each sample derived from an individual mouse brain (n = 12). (D) Representative immunoblots illustrating phospho-cPLA₂ (110 kDa) and β-actin (42 kDa) protein. Relative integrated density values of phospho-cPLA₂ in brain from COX-1^−/− mice are expressed as percentage of wild type. Each sample derived from an individual mouse brain (n = 12). *p ≤ 0.05, **p ≤ 0.01 versus wild type.