Figure 5
Figure 5. Immunophenotype of WT, PU/ER(T)+/−, and PU/ER(T)+/+ bone marrow chimera mice. (A) WT, PU/ER(T)+/−, and PU/ER(T)+/+ bone marrow chimera mice were injected with luciferin 1 mg/mL and imaged to measure bioluminescence which is an indirect indicator of NF-κB activation at 0 and 16 hours after LPS (10 mg/kg body weight) injection. NF-κB activation in terms of chest photon values per second per square centimeter area was quantitated (represented in circled chest area). Data represent mean ± SEM of 5 individual measurements for LPS treatment and controls (*P < .001 compared with WT LPS treatment, **P < .01 compared with WT LPS treatment). (B) Lung, liver, and spleen tissue from WT, PU/ER(T)+/−, and PU/ER(T)+/+ bone marrow chimera mice were collected, and homogenates were prepared in luciferase assay tissue lysis buffer (Promega). The luciferase activity was measured in different tissue homogenates using luciferin as the substrate. Data represent mean ± SD of 5 individual measurements for LPS and vehicle treatments (*P < .001 compared with WT LPS treatment, **P < .01 compared with WT LPS treatment). (C) Sixteen hours after LPS treatment, BAL fluid was collected from different groups of mice; cytospin slides were prepared and stained with HEMA 3. The numbers of macrophages and neutrophils per field view of microscope were quantitated. Data represent mean ± SD of 3 individual measurements from different animals (*P < .001 compared with WT LPS treatment, **P < .01 compared with WT LPS treatment). (D) Lung tissue homogenates from different groups of bone marrow chimera mice were prepared and analyzed for MPO activity. One unit of MPO activity was defined as change in absorbance at 460 nm per milligram of protein per minute. Data represent mean ± SD of 3 individual measurements from different animals (*P < .001 compared with WT LPS treatment, **P < .01 compared with WT LPS treatment). (E) Blood serum was collected from vehicle and LPS-treated bone marrow chimera mice, and levels of IL-6, MCP-1, KC, TNF-α, IL-1β, IL-10, IL-12p40, and IL-12p70 were measured by multiplex ELISA. Data represent mean ± SD of 4 individual measurements from different animals (*P < .001 compared with WT LPS treatment, **P < .01 compared with WT LPS treatment).

Immunophenotype of WT, PU/ER(T)+/−, and PU/ER(T)+/+ bone marrow chimera mice. (A) WT, PU/ER(T)+/−, and PU/ER(T)+/+ bone marrow chimera mice were injected with luciferin 1 mg/mL and imaged to measure bioluminescence which is an indirect indicator of NF-κB activation at 0 and 16 hours after LPS (10 mg/kg body weight) injection. NF-κB activation in terms of chest photon values per second per square centimeter area was quantitated (represented in circled chest area). Data represent mean ± SEM of 5 individual measurements for LPS treatment and controls (*P < .001 compared with WT LPS treatment, **P < .01 compared with WT LPS treatment). (B) Lung, liver, and spleen tissue from WT, PU/ER(T)+/−, and PU/ER(T)+/+ bone marrow chimera mice were collected, and homogenates were prepared in luciferase assay tissue lysis buffer (Promega). The luciferase activity was measured in different tissue homogenates using luciferin as the substrate. Data represent mean ± SD of 5 individual measurements for LPS and vehicle treatments (*P < .001 compared with WT LPS treatment, **P < .01 compared with WT LPS treatment). (C) Sixteen hours after LPS treatment, BAL fluid was collected from different groups of mice; cytospin slides were prepared and stained with HEMA 3. The numbers of macrophages and neutrophils per field view of microscope were quantitated. Data represent mean ± SD of 3 individual measurements from different animals (*P < .001 compared with WT LPS treatment, **P < .01 compared with WT LPS treatment). (D) Lung tissue homogenates from different groups of bone marrow chimera mice were prepared and analyzed for MPO activity. One unit of MPO activity was defined as change in absorbance at 460 nm per milligram of protein per minute. Data represent mean ± SD of 3 individual measurements from different animals (*P < .001 compared with WT LPS treatment, **P < .01 compared with WT LPS treatment). (E) Blood serum was collected from vehicle and LPS-treated bone marrow chimera mice, and levels of IL-6, MCP-1, KC, TNF-α, IL-1β, IL-10, IL-12p40, and IL-12p70 were measured by multiplex ELISA. Data represent mean ± SD of 4 individual measurements from different animals (*P < .001 compared with WT LPS treatment, **P < .01 compared with WT LPS treatment).

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