Journal: Cell cycle (Georgetown, Tex.)

Article Title: Irreversible chromosome damage accumulates rapidly in the absence of ATM kinase activity.

doi: 10.4161/cc.7.9.5961

Figure Lengend Snippet: Figure 1. (A) Detection of phosphorylation on ectopically expressed ATM. Constructs expressing Flag-tagged ATM wild-type kinase (WT), ATM kinase-dead (KD) or ATM S1981A (1981A) were immunoprecipitated from H460 cells 15 min following exposure to 5 Gy g-rays, and assessed by immunoblotting as indicated. The 1981S-P antisera recognizes WT- but not KD- or S1981A-ATM following irradiation. (B) Anti-1981S-P recognizes a single protein induced by IR in crude extracts prepared from exponentially growing primary fibroblasts. Whole-cell extract was prepared 15 min after 5 Gy g-rays, and samples were resolved in a 4–12% gradient gel and immunoblotted with anti-1981S-P antisera. (C) Sensitivity of anti-1981S-P antisera. IMR90 cells were lysed 15 min following increasing doses of irradiation, and the levels of ATM serine 1981 phosphorylation and total ATM were determined by immunoblotting.

Article Snippet: The transformed lung cell line NCI-H460, which is hypotriploid but wild-type for p53, and the normal diploid fetal human lung fibroblast cell line IMR90 were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Phospho-proteomics, Construct, Expressing, Immunoprecipitation, Western Blot, Irradiation

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Irreversible chromosome damage accumulates rapidly in the absence of ATM kinase activity.

doi: 10.4161/cc.7.9.5961

Figure Lengend Snippet: Figure 4. The frequency of chromosome aberrations is increased 48 h following irradiation in KU55933 treated late-S and G2 cells. Cells were treated with 10 μM KU55933 from +15 min to +75 min following exposure to 2 Gy g-rays. Cells were harvested 48 h later following an exposure to 250 nM colcemid (Irvine Scientific), a microtubule inhibitor, or 50 nM calyculin A (Calbiochem), which prematurely condenses chromatin, allowing visualization of late-S- and G2-phase cells. In order to compensate for differences in ploidy between cell lines, it is more appropriate to compare the total weighted aberrations per cell. In both H460 (A) and IMR90 (B) cells, there was ~1 break per cell, irrespective of KU55933 post-treatment or irradiation, with the exception of the cells exposed to both KU55933 and IR. H460 and IMR90 cells exposed to both KU55933 and IR contained at least a two-fold increase in aberrations per cell.

Article Snippet: The transformed lung cell line NCI-H460, which is hypotriploid but wild-type for p53, and the normal diploid fetal human lung fibroblast cell line IMR90 were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Irradiation

Journal: Prostaglandins, leukotrienes, and essential fatty acids

Article Title: Prostaglandin E₂ possesses different potencies in inducing Vascular Endothelial Growth Factor and Interleukin-8 production in COPD human lung fibroblasts.

doi: 10.1016/j.plefa.2016.01.005

Figure Lengend Snippet: Fig. 1. PGE2 (Panel A), VEGF (Panel B), and IL-8 (Panel C) concentrations in supernatants of cultured lung fibroblasts from control subjects (n¼6), smoker (n¼6) and COPD patients (n¼8). Box plot represents median and IQ range. Statistical analysis was performed by ANOVA and Fisher’s PLSD correction.

Article Snippet: Human fetal lung fibroblasts (HFL-1; lung, diploid, human) (from American Type Culture Collection, Manassas V.A.) were cultured in 100 mm tissue culture dishes (Falcon; BectonDickinson Labware, Lincoln Park, NJ) with RPMI1640 supplemented with 10% FCS, 2 mM glutamine, 25 mM HEPES buffer, 1% MEM non essential aminoacids, 100 U/ml penicillin, 100 mg/ml streptomycin, and 2.5 μg/ml fungizone.

Techniques: Cell Culture, Control

Journal: Prostaglandins, leukotrienes, and essential fatty acids

Article Title: Prostaglandin E₂ possesses different potencies in inducing Vascular Endothelial Growth Factor and Interleukin-8 production in COPD human lung fibroblasts.

doi: 10.1016/j.plefa.2016.01.005

Figure Lengend Snippet: Fig. 2. COX-1, COX-2, EP1, EP2, EP3, and EP4 protein expression in cultured lung fibroblasts from control subjects (n¼6), smoker (n¼6) and COPD patients (n¼8). The results were expresses in arbitrary densitometric units (ADU) corrected against the density of β-actin bands. Panel A: COX-1 (top panel) and COX-2 (middle panel) and repre- sentative gel images of COX-1 and COX-2 (bottom panel). Panel B: EP1 and EP2 (top panels), EP3 and EP4 (middle panels) and representative gel images of EP1, EP2, EP3, and EP4 (bottom panel). Statistical analysis was performed by ANOVA and Fisher’s PLSD correction.

Article Snippet: Human fetal lung fibroblasts (HFL-1; lung, diploid, human) (from American Type Culture Collection, Manassas V.A.) were cultured in 100 mm tissue culture dishes (Falcon; BectonDickinson Labware, Lincoln Park, NJ) with RPMI1640 supplemented with 10% FCS, 2 mM glutamine, 25 mM HEPES buffer, 1% MEM non essential aminoacids, 100 U/ml penicillin, 100 mg/ml streptomycin, and 2.5 μg/ml fungizone.

Techniques: Expressing, Cell Culture, Control

Journal: Prostaglandins, leukotrienes, and essential fatty acids

Article Title: Prostaglandin E₂ possesses different potencies in inducing Vascular Endothelial Growth Factor and Interleukin-8 production in COPD human lung fibroblasts.

doi: 10.1016/j.plefa.2016.01.005

Figure Lengend Snippet: Fig. 3. : COX2, EP2, EP4 gene expression in cultured lung fibroblasts from control subjects (n¼3), smoker (n¼5) and COPD patients (n¼6). Data are expressed as fold increase versus the control group that was chosen as the reference group (see Section 2 for details). Statistical analysis was performed by Kruskal–Wallis test and Mann- Whitney U test.

Article Snippet: Human fetal lung fibroblasts (HFL-1; lung, diploid, human) (from American Type Culture Collection, Manassas V.A.) were cultured in 100 mm tissue culture dishes (Falcon; BectonDickinson Labware, Lincoln Park, NJ) with RPMI1640 supplemented with 10% FCS, 2 mM glutamine, 25 mM HEPES buffer, 1% MEM non essential aminoacids, 100 U/ml penicillin, 100 mg/ml streptomycin, and 2.5 μg/ml fungizone.

Techniques: Gene Expression, Cell Culture, Control, MANN-WHITNEY

Journal: Prostaglandins, leukotrienes, and essential fatty acids

Article Title: Prostaglandin E₂ possesses different potencies in inducing Vascular Endothelial Growth Factor and Interleukin-8 production in COPD human lung fibroblasts.

doi: 10.1016/j.plefa.2016.01.005

Figure Lengend Snippet: Fig. 4. : PGE2 stimulates VEGF and IL-8 release in human fetal lung fibroblasts (HFL-1) in a dose-dependent manner. Cultured HFL-1 were stimulated with different concentrations of PGE2 (109 M–106 Μ) for 18 h. At the end of stimulation, cell supernatants were collected for VEGF (Panel A) and IL-8 (Panel B) analysis by ELISA (See Methods for details). The results were expressed as mean7SD from four independent experiments. Statistical analysis was performed by ANOVA and Fisher’s PLSD cor- rection. *po0.05, **po0.01, and ***po0.001 vs basal;.

Article Snippet: Human fetal lung fibroblasts (HFL-1; lung, diploid, human) (from American Type Culture Collection, Manassas V.A.) were cultured in 100 mm tissue culture dishes (Falcon; BectonDickinson Labware, Lincoln Park, NJ) with RPMI1640 supplemented with 10% FCS, 2 mM glutamine, 25 mM HEPES buffer, 1% MEM non essential aminoacids, 100 U/ml penicillin, 100 mg/ml streptomycin, and 2.5 μg/ml fungizone.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Prostaglandins, leukotrienes, and essential fatty acids

Article Title: Prostaglandin E₂ possesses different potencies in inducing Vascular Endothelial Growth Factor and Interleukin-8 production in COPD human lung fibroblasts.

doi: 10.1016/j.plefa.2016.01.005

Figure Lengend Snippet: Fig. 5. VEGF and IL-8 production from human fetal lung fibroblasts (HFL-1) stimulated with supernatants of lung fibroblasts with low PGE2 level (o109 M, n¼3), and with high PGE2 level (4108 M, n¼3), (see Methods for details). Values are expressed as mean7SD Panel A: VEGF (pg/ml) (top) and IL-8 (pg/ml) (bottom) production after stimulation with and without supernatants of lung fibroblasts with low PGE2 level. Panel B: VEGF (pg/ml) (top) and IL-8 (pg/ml) (bottom) production after stimulation with and without supernatants of lung fibroblasts with high PGE2 level and with supernatants depleted from PGE2. Statistical analysis was performed by ANOVA and Fisher’s PLSD correction.

Article Snippet: Human fetal lung fibroblasts (HFL-1; lung, diploid, human) (from American Type Culture Collection, Manassas V.A.) were cultured in 100 mm tissue culture dishes (Falcon; BectonDickinson Labware, Lincoln Park, NJ) with RPMI1640 supplemented with 10% FCS, 2 mM glutamine, 25 mM HEPES buffer, 1% MEM non essential aminoacids, 100 U/ml penicillin, 100 mg/ml streptomycin, and 2.5 μg/ml fungizone.

Techniques:

Journal: Prostaglandins, leukotrienes, and essential fatty acids

Article Title: Prostaglandin E₂ possesses different potencies in inducing Vascular Endothelial Growth Factor and Interleukin-8 production in COPD human lung fibroblasts.

doi: 10.1016/j.plefa.2016.01.005

Figure Lengend Snippet: Fig. 6. Effect of specific EP receptors agonists and antagonists on VEGF and IL-8 release from human fetal lung fibroblasts (HFL-1). HFL-1 cells were stimulated with the indicated agonists or with the supernatants of lung fibroblasts from COPD (see Methods for details). Values are expressed as mean7SD. Production of VEGF (Panel A) and IL-8 (Panel B) in the presence or absence of Butaprost (EP2 agonist) or Sulprostone (EP3 agonist). Panel C: Production of VEGF after stimulation with supernatants of lung fibroblasts from COPD in presence or absence of SC-19220 (EP1 antagonist) or AH-23848 (EP4 antagonist). Statistical analysis was performed by ANOVA and Fisher’s PLSD correction.

Article Snippet: Human fetal lung fibroblasts (HFL-1; lung, diploid, human) (from American Type Culture Collection, Manassas V.A.) were cultured in 100 mm tissue culture dishes (Falcon; BectonDickinson Labware, Lincoln Park, NJ) with RPMI1640 supplemented with 10% FCS, 2 mM glutamine, 25 mM HEPES buffer, 1% MEM non essential aminoacids, 100 U/ml penicillin, 100 mg/ml streptomycin, and 2.5 μg/ml fungizone.

Techniques: