Display Settings:

Items per page

Results: 8

1.
Figure 6

Figure 6. From: Enhanced Toll-like receptor responses in the absence of signaling adaptor DAP12.

LPS-induced ERK phosphorylation is increased in DAP12-deficient macrophages. Bone marrow-derived macrophages from wild-type or DAP12-deficient (DAP12 KO) mice were stimulated with 0.5 ng/ml LPS for the indicated time (min), at which time cells were lysed. Cytoplasmic extracts were analyzed using antibodies specific for p38 MAPK, p42/44 ERK and JNK, or for phosphorylated versions of these proteins. Data are representative of four independent experiments.

Jessica A Hamerman, et al. Nat Immunol. ;6(6):579-586.
2.
Figure 1

Figure 1. From: Enhanced Toll-like receptor responses in the absence of signaling adaptor DAP12.

DAP12-deficient macrophages secrete increased amounts of cytokines after TLR stimulation. Bone marrow-derived macrophages from wild-type or DAP12-deficient (DAP12 KO) mice were incubated with the indicated concentrations of LPS, CpG DNA, synthetic bacterial lipopeptide or zymosan for 16 h. Supernatants were collected and the amounts of TNF (a), IL-6 (b) and IL-12 p40 (c) were measured. No IL-6 was detected after incubation of macrophages with zymosan. Data are representative of ten (LPS, zymosan; a), five (lipopeptide, CpG DNA; a) or four (b,c) independent experiments.

Jessica A Hamerman, et al. Nat Immunol. ;6(6):579-586.
3.
Figure 4

Figure 4. From: Enhanced Toll-like receptor responses in the absence of signaling adaptor DAP12.

Syk-deficient macrophages secrete increased amounts of proinflammatory cytokines in response to TLR stimulation. Bone marrow-derived macrophages from wild-type mice, DAP12-deficient (DAP12 KO) mice or Syk-deficient (Syk KO) fetal liver chimeras were incubated with the indicated concentrations of LPS, CpG DNA, or synthetic bacterial lipopeptide for 16 h. Supernatants were collected and the amounts of TNF (a), IL-6 (b) and IL-12 p40 (c) were measured using ELISA. Data are representative of three independent experiments.

Jessica A Hamerman, et al. Nat Immunol. ;6(6):579-586.
4.
Figure 8

Figure 8. From: Enhanced Toll-like receptor responses in the absence of signaling adaptor DAP12.

DAP12-deficient mice have an enhanced innate immune response to infection with L. monocytogenes. Wild-type and DAP12-deficient mice were infected intravenously with 5 × 104 (a) or 1 × 104 (b) CFU of L. monocytogenes strain 10403S. Three days after infection, the number of CFU in the spleens and livers of the mice were determined. The data are represented as the CFU in each individual mouse (circles) and the mean (horizontal line). In a, there were five mice per group; in b there were six wild-type mice and four DAP12-deficient mice per group. These data are representative of four independent experiments.

Jessica A Hamerman, et al. Nat Immunol. ;6(6):579-586.
5.
Figure 7

Figure 7. From: Enhanced Toll-like receptor responses in the absence of signaling adaptor DAP12.

DAP12-deficient mice are more susceptible to endotoxic shock than wild-type mice. Wild-type (n = 10) or DAP12-deficient (DAP12 KO; n = 9) mice were injected intraperitoneally with LPS and D-galactosamine and then monitored for survival for 12 h (a). Data are representative of four independent experiments. Wild-type (n = 5) and DAP12-deficient (n = 5) mice were injected with 100 μg of LPS. At the indicated times, a sample of blood was drawn and plasma was analyzed for TNF using ELISA (b). The data are reported as mean ± s.e.m. and are representative of three independent experiments. Mice were treated as described in b and plasma TNF concentrations at 1 h postinjection are represented as the mean of 25 mice assayed in five independent experiments ± s.e.m (c). The difference between wild-type and DAP12-deficient mice is significant using Student’s t-test, P < 0.03 (two-tailed distribution).

Jessica A Hamerman, et al. Nat Immunol. ;6(6):579-586.
6.
Figure 3

Figure 3. From: Enhanced Toll-like receptor responses in the absence of signaling adaptor DAP12.

Reintroduction of DAP12 reduces the TNF production by DAP12-deficient macrophages in an ITAM-dependent manner. Bone marrow-derived macrophages from wild-type or DAP12-deficient (DAP12 KO) mice were transduced with an empty vector (control) retrovirus or retroviruses encoding wild-type DAP12 or a DAP12 ITAM mutant (DAP12Y→F). The macrophages were activated with zymosan (ten per macrophage) or CpG DNA (0.01 μM) in the presence of Brefeldin A for 4 h and TNF production was assessed by flow cytometry. Transduced cells were gated based on GFP fluorescence and the percentage of TNF-producing cells was determined. (a) Histograms of TNF expression in wild-type and DAP12-deficient macrophages transduced with control or DAP12-expressing retrovirus from one representative experiment. (b,c) The percentage of transduced cells producing TNF represented as the mean ± s.e.m. of three independent experiments (zymosan) or two independent experiments (CpG DNA).

Jessica A Hamerman, et al. Nat Immunol. ;6(6):579-586.
7.
Figure 2

Figure 2. From: Enhanced Toll-like receptor responses in the absence of signaling adaptor DAP12.

DAP12-deficient macrophages produce more TNF after TLR stimulation. (a,b) Bone marrow–derived macrophages from wild-type or DAP12-deficient (DAP12 KO) mice were incubated with LPS (0.4 ng/ml), CpG DNA (0.04 μM) or zymosan (ten particles per macrophage) for 4 h (CpG DNA and zymosan) or 14 h (LPS) in the presence of Brefeldin A for the final 4 h. TNF secretion was then assessed; data are represented by one-parameter histograms (a) or by the mean fluorescent intensity (MFI) of the TNF-producing population (b). Data are representative of four independent experiments. (c) Bone marrow–derived macrophages (day 7) were stimulated with LPS (0.5 ng/ml) or zymosan (ten particles per macrophage). Supernatants were removed at the indicated time points, and TNF concentrations were measured using ELISA. Data are representative of three independent experiments.

Jessica A Hamerman, et al. Nat Immunol. ;6(6):579-586.
8.
Figure 5

Figure 5. From: Enhanced Toll-like receptor responses in the absence of signaling adaptor DAP12.

Soluble inhibitory factor production does not explain differences between wild-type and DAP12-deficient macrophages. (a) Bone marrow-derived macrophages from wild-type and DAP12-deficient (DAP12 KO) mice were stimulated with the indicated concentrations of LPS, zymosan, CpG DNA and lipopeptide for 16 h. Supernatants were then assayed for TNF or IL-10 using ELISA. (b) Wild-type and DAP12-deficient macrophages were cultured in the top and bottom chambers of culture plates separated by a porous membrane. Cells in both top and bottom chambers were treated with CpG DNA (0.01 μM) or zymosan (ten particles per macrophage) for 4 h, and TNF-producing cells in the bottom chambers were determined by using flow cytometry (). In the histograms shown, the type of cells in the top and bottom chambers of the culture plates are indicated as cells in bottom chamber:cells in top chamber. The percentage of TNF-positive cells is indicated in the top right corner of each histogram. Data shown are representative of two independent experiments.

Jessica A Hamerman, et al. Nat Immunol. ;6(6):579-586.

Display Settings:

Items per page

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk