STAT-6 phosphorylation following injection with IL-13.

<p>Positive control (PC) consists of lung nuclear proteins from IL4Ra+/− embryos directly injected with IL-13. Negative controls consist of lung nuclear protein from IL4Ra−/− females injected with IL-13 (NC1) and nuclear protein from IL4Ra+/− embryos injected with PBS (NC2). The experimental group consists of nuclear proteins from IL4Ra+/− embryos whose mothers were injected with IL-13. Bands for P-STAT-6 are only detectable in PC lane. HDAC1 was used as a loading control. Insufficient protein was loaded in NC1, but otherwise protein loading was similar.</p

Growth rate of the continuous murine alveolar macrophage single cell clones ZK1 (◆), ZK2 (■) and ZK6 (▲)

Cells were seeded in six-well plate at 2 × 10cells/ml and incubated at 37°C in a 5% CO-humidified atmosphere in RPMI/10% FBS complete medium. Three wells per clone were harvested with cold PBS at 14 h and 18 h post seeding, and cells from each well were counted by a hemocytometer with trypan blue exclusion of dead cells. The obtained average cell count for each clone at each time point was plotted against the time. Doubling time (mean generation time = mgt) was calculated according the formula: N = N2. N is the number of cells at any time T; N, is the number of cells at an initial point. The doubling time of ZK cell lines is approximately 14 hours.<p><b>Copyright information:</b></p><p>Taken from "Characterization of immortalized MARCO and SR-AI/II-deficient murine alveolar macrophage cell lines"</p><p>http://www.particleandfibretoxicology.com/content/5/1/7</p><p>Particle and Fibre Toxicology 2008;5():7-7.</p><p>Published online 2 May 2008</p><p>PMCID:PMC2427050.</p><p></p

ZK1, ZK2 and ZK6 cell lines are MARCOand SR-AI/II(MS) by PCR genotyping

With primers for SR-A, amplifies a 325 bp DNA fragment from the C57BL/6 wild-type (WT) allele; with SR-AI/II mutant allele primers, amplifies a 434 bp DNA fragment from SRA-deficient ZK1, ZK2 and ZK6 cells. With primers for MARCO wild-type allele, amplifies a 500 bp DNA fragment from WT mice; with primers for MARCO mutant allele, amplifies a 850 bp DNA fragment from ZK cells. ZK1, ZK2 and ZK6 clones exhibited both MARCO and SRA-I/II-deficient. PCR products, ca.10 μl/each was resolved on a 1.5% agarose gel by gel electrophoresis. M, 100 bp DNA marker.<p><b>Copyright information:</b></p><p>Taken from "Characterization of immortalized MARCO and SR-AI/II-deficient murine alveolar macrophage cell lines"</p><p>http://www.particleandfibretoxicology.com/content/5/1/7</p><p>Particle and Fibre Toxicology 2008;5():7-7.</p><p>Published online 2 May 2008</p><p>PMCID:PMC2427050.</p><p></p

ZK cell lines and primary AMs with MS significantly diminished phagocytosis of fluorescent latex beads

Wild type primary AMs were used as control. Values shown are the means ± SD from four separate experiments. **Significant difference from WT control, < 0.001. () Polyinosinic acid (PolyI) had no effect but dextran sulfate (DS) increased binding by ZK1 cells. In contrast, PolyI and DS marked reduced wild-type AM binding of the latex beads. Data are expressed as the mean ± SD and compared to the control in each group (WT, MS, ZK1, respectively). *Significant difference compared with ZK1 (< 0.05); **Significant difference compared with WT (< 0.001). PolyI and DS (500 kDa), 10 μg/ml each. () The inhibition of ZK1 cells binding of the latex beads by dextran sulfate was size-dependent. Only smaller 5-8-kDa dextran sulfate was able to inhibit ZK1 cells binding of the latex beads. Data were shown as means ± SD. * < 0.05 versus control (n = 4). DS, 10 μg/ml.<p><b>Copyright information:</b></p><p>Taken from "Characterization of immortalized MARCO and SR-AI/II-deficient murine alveolar macrophage cell lines"</p><p>http://www.particleandfibretoxicology.com/content/5/1/7</p><p>Particle and Fibre Toxicology 2008;5():7-7.</p><p>Published online 2 May 2008</p><p>PMCID:PMC2427050.</p><p></p

Fc-receptor on ZK1 cells mediated phagocytosis of opsonized sheep red blood cells (SRBC)

ZK1 cells were plated at 1 × 10cells/well in a 6-well plate containing a sterile micro cover glass per well in RPMI complete medium for overnight at 37°C. Unopsonized (as negative control) or preopsonized SRBC were plated on monolayer of ZK1 cells at a ratio of 20:1 and incubated at 37°C for 1 h. After removal of free SRBC by medium exchange and lysis by osmotic shock, the cells on the cover glass were fixed and stained with a modified Wright stain, subsequently examined by light microscopy. Panel , ZK1 cells were unable to ingest unopsonized SRBC after lysis. Some free SRBC were present without lysis as background. Panel , approximately 80% of ZK1 cells were positive phagocytosis of opsonized SRBC.<p><b>Copyright information:</b></p><p>Taken from "Characterization of immortalized MARCO and SR-AI/II-deficient murine alveolar macrophage cell lines"</p><p>http://www.particleandfibretoxicology.com/content/5/1/7</p><p>Particle and Fibre Toxicology 2008;5():7-7.</p><p>Published online 2 May 2008</p><p>PMCID:PMC2427050.</p><p></p

Modified Wright staining of ZK1 and ZK2 cell lines compared with primary alveolar macrophages

Primary AMs were isolated from wild type (WT) C57BL/6 mice and from MARCOand SR-AI/II(MS) mice. ZK1 and ZK2 cells were identified as macrophages by their large, dark nuclei and abundant pale, granular cytoplasm containing numerous vacuoles.<p><b>Copyright information:</b></p><p>Taken from "Characterization of immortalized MARCO and SR-AI/II-deficient murine alveolar macrophage cell lines"</p><p>http://www.particleandfibretoxicology.com/content/5/1/7</p><p>Particle and Fibre Toxicology 2008;5():7-7.</p><p>Published online 2 May 2008</p><p>PMCID:PMC2427050.</p><p></p

In the allergen stimulated group DCs most genes with significant change in transcription (DE) had altered methylation either in the promoter (14%), CpG island (22%) or both (38%) (of all overalpping DMR-DE).

<p>Only 26% of the genes had changes elsewhere in the body of the transcript.</p

Scanning cytometry fluorescence imaging with GFP-S. aureus RN6390.

<p>Adherent GM-MФs were incubated with unopsonized GFP-<i>S. aureus</i> RN6390. External <i>S. aureus</i> were labeled with an IgG₃ monoclonal mouse anti-<i>S. aureus</i> primary antibody and Texas-RedX-conjugated goat anti-mouse secondary antibody. Collapsed confocal stack images were acquired by scanning cytometry. (A) CellTracker Blue and Hoechst channel (cells). (B) GFP channel (all bacteria). (C) Texas Red channel (external bacteria). (D) Composite image. (Original magnification, 200X).</p

Summary of binding inhibition results.

<p>Beads: biotinlyated carboxylated 1 micron green fluorescent latex beads. CK Wood: commercial killed AlexaFluor 488-conjugated <i>S. aureus</i> Wood strain.</p><p>Wood: live S. aureus Wood strain. Seattle 1945: Live S. aureus Seattle 1945 strain. RN 6390: live GFP+S. aureus RN6390 strain. IgG ops. RN6390: IgG α-S. aureus opsonized RN6390.</p><p>+ <25% inhibition. ++25–50% inhibition. +++50–75% inhibition. ++++ >75% inhibition.</p><p>- not significantly changed. ̂̂ 25–50% increased. nd: not determined.</p

Number of transcripts differentially methylated and differentially expressed between asthma-at-risk DCs and controls.

<p>Number of transcripts differentially methylated and differentially expressed between asthma-at-risk DCs and controls.</p