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Line was purchased from ATCC, and cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10 fetal bovine serum (FBS) and gentamycin (50 g/ml). ZK cell lines in this study were cultured in RPMI 1640 complete medium, in which RPMI medium was supplemented with 10 FBS, L-glutamine (2 mM), penicillin (100 U/ml), streptomycin (100 g/ ml). Isolation of alveolar macrophages Alveolar mac
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L antibodies made also rapidly clear to the clinicians that a reliable predictive factor for outcome was, in fact, lacking [3-7]. The introduction of K-RAS mutational status analysis allowed a reliable selection of resistant patients (i.e. those with mutated K-RAS). However not all K-RAS wildtype cases were also responders to anti-EGFR monoclonal antibodies. This observation made the need for furt
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Hat three clones ZK1, ZK2 and ZK6 were obtained by limiting dilution and further characterized. Our PCR genotyping results verified these three clones, ZK1, ZK2 and ZK6 are MARCO-/- and SR-AI/ II-/--deficient (Fig. 1). These cell lines are able to grow rapidly in RPMI or DMEM complete media in the absence of exogenous M-CSF or other growth factors, and their doubling time is 12?6 h on the average
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Ure Collection (ATCC, Manassas, VA); RPMI 1640 medium and PBS were purchased from BioWhittaker (Walkersville, MD); Gentamycin, penicillin/streptomycin, macrophage-colony stimulating factor (M-CSF), and Polybrene were obtained from Sigma (St. Louis, MO); MethoCultTM GF M3434, semi-solid medium was obtained from StemCell Technologies (Vancouver, Canada); DiffQuik, a modified Wright-Giemsa stain was
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C capacity functionally, but like primary AMs deficient in MARCO and SR-AI/II, ZK cells were significantly impaired in phagocytosis compared to the WT AMs due to deficiency in scavenger receptors.-Page 5 of(page number not for citation purposes)Particle and Fibre Toxicology 2008, 5:http://www.particleandfibretoxicology.com/content/5/1/A. Unopsonized Red Blood CellsNo lysisLysisB. Opsonized Red Blo
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Ure Collection (ATCC, Manassas, VA); RPMI 1640 medium and PBS were purchased from BioWhittaker (Walkersville, MD); Gentamycin, penicillin/streptomycin, macrophage-colony stimulating factor (M-CSF), and Polybrene were obtained from Sigma (St. Louis, MO); MethoCultTM GF M3434, semi-solid medium was obtained from StemCell Technologies (Vancouver, Canada); DiffQuik, a modified Wright-Giemsa stain was
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Surface expression of the macrophage-associated differentiation Ags was assessed by direct immunofluorescence (thick lines). ZK1 cells were incubated with FITC-labeled anti-mouse F4/80 or FITC-labeled anti-mouse CD11b. Mouse IgG2a and IgG2b were used as isotype controls (light lines). Staining of cells with FITC-labeled anti-mouse Ig compared to unstained cells detects the of cells expressing su
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S from MARCO-/- and SR-AI/II-/- mice, they demonstrate marked decreased binding and phagocytosis of these particles compared to primary AMs from wild type mice (Fig. 6A and Fig. 7A and Fig. 7B). The results confirm that MARCO and SR-AI/II receptors on alveolar macrophages play critical roles in uptake of unopsonized environmental particles and bacteria. Moreover, the reduced, but still demonstrabl