Figure 6.

Genetic blockade of PLCβ2/β3 signalling inhibits fMLP activation of Ras, p42/p44 MAPK, PKB and PIP₃ accumulation. Neutrophils were prepared from either PLCβ2^−/− × PLCβ3^−/− or PLCβ2^+/+ × PLCβ3^+/+ mice, stimulated with fMLP (10 μM for 1 min), or its vehicle, and a range of responses were quantified. (A) Lower part shows representative immunoblots of Ras activation by pull down, and phosphorylation of S473-PKB (P-PKB) and T202/Y204-p42/p44 MAPK (P-p42/p44-MAPK) and total Ras (as a loading control) in aliquots of the lysates from which the pull downs were performed. The upper part shows quantification of data, like that below, from three independent experiments and are expressed as means±s.e., n=8, as a % of the stimulated samples in that experiment, normalized via their respective loading controls. (B, C) Measurements of neutrophil stearoyl/arachidonoyl-DAG or stearoyl/arachidonoyl-PIP₃ as response ratios with the ISD recovered in that sample are shown. The data presented are means±s.d., n=8, from three independent experiments. Figure source data can be found with the Supplementary data.