Fig. 1. Schematic diagram of mutations created in PI3Kγ subunits and their effect on expression levels in isolated bone marrow neutrophils (BMN).

(A) Cartoon illustrating Gβγ binding to p84-p110γ and p101-p110γ complexes and the mutations created in the Gβγ binding domains of p101 regulatory and p110γ catalytic subunits. A single Gβγ binding site is depicted in p101-p110γ but it is currently unknown whether there is a single or multiple binding sites for Gβγ in this complex. (B to E) Relative abundance of PI3Kγ subunits in BMN derived from the indicated strains of mice; (B) Western blot representative of five independent experiments that were combined and quantified for the abundance of p84 (C), p110γ (D) and p101 (E). Western blotting analysis was performed on 1x10⁶ BMNs from mice of the indicated genotypes. The abundances of p84, p110γ and p101 protein in samples from mice of each genotype were quantitated by densitometry and normalized to p67^phox to correct for neutrophil input (β-COP is shown as an additional loading control). Data are means ± SEM of n= 5 independent experiments and are expressed as a ratio to the relative protein band density of WT samples on each blot. Normalized data was analyzed by ratio paired t tests with Holms-Sidak corrections applied to account for multiple comparisons, with comparisons made to WT samples.