Fig. 7. Expression of mutant cPLA₂α reproduces the KI phenotype in KO pDFs.

(A) Immunoblotting and quantification of cPLA₂α in pDFs from WT, cPLA₂α KI, and cPLA₂α KO mice. β-actin is a loading control. n = 3 cell isolates per genotype (3–4 mice per genotype were utilized to generate the cell isolates). (B) Immunoblotting for cPLA₂α in cPLA₂α KO pDFs in which an adenoviral (CMV) expression system (30) was used to drive expression of WT cPLA₂α or C1P-binding mutant cPLA₂α. Empty vector is a negative control. Data is representative of n = 3 independent experiments using cells isolated from different cPLA₂α KO mice. (C) Lipid profiles of WT and KI pDFs expressing WT or mutant cPLA₂α from the CMV vector. (D) Quantification of 5-HETE in KO pDFs expressing WT or mutant cPLA₂α from the CMV vector. For (C) and (D), n = 3 (empty CMV control) and n = 9 (WT and mutant cPLA₂α) cell isolates from 5 cPLA₂α KO mice. (E) Migration velocity in KO pDFs expressing WT or mutant cPLA₂α from the CMV vector. n = 12 cell isolates per genotype (6–9 mice per genotype were used to generate the cell isolates). Samples were compared using ANOVA followed by Tukey’s post-hoc test. Data shown are means ± SD*P< 0.05, **P< 0.01, ***P< 0.001, ****P< 0.0001.