Analysis of ER stress, apoptosis, and functionality in G6pc3^−/− macrophages. Peritoneal macrophages were isolated from 6- to 8-week-old wild-type (+/+) and G6pc3^−/− (−/−) littermates. Freshly isolated macrophages were used to examine ER stress and apoptosis. For functional analysis, macrophages were further enriched by CD11b microbeads binding. (A) Western blot analysis of protein extracts of macrophages with the use of Abs against GRP170, GRP78/GRP94, or β-actin. Data from 2 pairs of littermates are shown, and each lane contains 50 μg of protein. (B) Quantification of apoptotic cells in macrophages of control (n = 6) and G6pc3^−/− (n = 6) mice during culturing in vitro in the absence or presence of BFA. Results represent the mean ± SEM. (C) Quantification of active caspase-3 in macrophages of control (n = 4) and G6pc3^−/− (n = 4) mice during culturing in vitro in the absence or presence of BFA. (D) The viability of CD11b-enriched macrophages from wild-type (n = 10) and G6pc3^−/− (n = 10) mice, estimated by flow cytometry. Results represent the mean ± SEM. (E) Macrophage respiratory burst activity in response to 200 ng/mL PMA. (F) Macrophage calcium flux in response to 10⁻⁷M of leukotriene D4. (G) Macrophage concentration-dependent chemotaxis in response to MCP-1 or M-CSF. (H) Macrophage phagocytosis activity. Quantification of bioparticle-positive macrophages in control and G6pc3^−/− mice; the numbers reflect the percentage of total macrophage that have engulfed particles. Data represent the mean ± SEM of 3 independent experiments. **P < .005 and *P < .05.