western blotting  (R&D Systems)

Journal: PLoS ONE

Article Title: Leukemia Inhibitory Factor (LIF) Inhibition during Mid-Gestation Impairs Trophoblast Invasion and Spiral Artery Remodelling during Pregnancy in Mice

doi: 10.1371/journal.pone.0129110

Figure Lengend Snippet: Primer sequences.

Article Snippet: Primary antibody for LIFRα (1:100; R&D Systems #AF-249NA), LIF (1:100; R&D Systems #AF-250NA), pan-cytokeratin (1:200; Santa Cruz #H-240), α-SMA (1:200; Dako #M085129), F4/80 (1:200; Serotec) or isotype negative control goat IgG in blocking solution were applied for 18h incubated at 4°C.

Techniques: Sequencing

Journal: PLoS ONE

Article Title: Leukemia Inhibitory Factor (LIF) Inhibition during Mid-Gestation Impairs Trophoblast Invasion and Spiral Artery Remodelling during Pregnancy in Mice

doi: 10.1371/journal.pone.0129110

Figure Lengend Snippet: Wild type (WT) mouse implantation sites were collected from n = 3 mice/time point. E6, 8 and 10 embryos were analyzed as whole implantation sites (IS) and E13, 15 and 17 were dissected to obtain the decidua only. (a) LIF, (b) LIFRα and (c) gp130 mRNA expression was determined by semi-quantitative PCR normalized to β2-microglobulin. Data are mean ± SEM, ANOVA, Tukey’s post-hoc test, *p<0.05, n = 3/time point.

Article Snippet: Primary antibody for LIFRα (1:100; R&D Systems #AF-249NA), LIF (1:100; R&D Systems #AF-250NA), pan-cytokeratin (1:200; Santa Cruz #H-240), α-SMA (1:200; Dako #M085129), F4/80 (1:200; Serotec) or isotype negative control goat IgG in blocking solution were applied for 18h incubated at 4°C.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Leukemia Inhibitory Factor (LIF) Inhibition during Mid-Gestation Impairs Trophoblast Invasion and Spiral Artery Remodelling during Pregnancy in Mice

doi: 10.1371/journal.pone.0129110

Figure Lengend Snippet: Wild type (WT) mouse implantation sites were collected from n = 3 mice/time point and 2μm serial sections were immunostained for cytokeratin, LIF and LIFRα. Representative photomicrographs of mid-gestation (E10 and E13) implantation site sections are shown here. Both LIF and LIFRα co-localized with cytokeratin in maternal decidual vessels. Bars represent 20μm. Insets are negative controls.

Article Snippet: Primary antibody for LIFRα (1:100; R&D Systems #AF-249NA), LIF (1:100; R&D Systems #AF-250NA), pan-cytokeratin (1:200; Santa Cruz #H-240), α-SMA (1:200; Dako #M085129), F4/80 (1:200; Serotec) or isotype negative control goat IgG in blocking solution were applied for 18h incubated at 4°C.

Techniques:

Journal: PLoS ONE

Article Title: The Phosphoinositide 3-Kinase p110α Isoform Regulates Leukemia Inhibitory Factor Receptor Expression via c-Myc and miR-125b to Promote Cell Proliferation in Medulloblastoma

doi: 10.1371/journal.pone.0123958

Figure Lengend Snippet: (a) LIFRα protein levels in medulloblastoma cell lines and normal brain (NB) tissue were evaluated by Western blot and compared with β-actin as the loading control. (b) LIF and LIFR expression in medulloblastoma cell lines was analyzed by semi quantitative RT-PCR. (c) Expression of PIK3CA and LIFR derived from the transcriptomic analysis of primary medulloblastoma, grouped according to molecular disease variants. Data from Northcott et al are shown. Gene expression levels were compared using the Kruskal-Wallis test and the p-value is shown at the bottom right of the illustration. As this global test was significant, Dunn's post hoc tests were used to compare the groups. P-values < 0,05, as determined by Dunn's post hoc tests, are indicated in order to illustrate the differences. (d) Analysis of cDNA microarray data performed on PTCH1 +/- ; TP53 +/- or PTCH1 +/- ; TP53 +/+ mice. The table represents a summary of fold change (logFC), average expression values (AveExpr), p values obtained by comparison of the two murine strains from Data set GEO accession number GSE37316). (e) Cell lysates of DAOY cells stimulated with or not with 5, 10, 25, 50 and 100 ng/ml of LIF for 10 min were analyzed for expression and phosphorylation of AKT and ERK pathway downstream targets. Treatment of 10 ng/ml of OSM for 10 min was used as a positive control.

Article Snippet: The Anti-human LIFRα antibody (AF-249-NA) and the recombinant rh-LIF Rα (7487-LR) were purchased from R&D Systems (Minneapolis, MN, USA) and were diluted directly into the medium immediately before use.

Techniques: Western Blot, Control, Expressing, Quantitative RT-PCR, Derivative Assay, Gene Expression, Microarray, Comparison, Phospho-proteomics, Positive Control

Journal: PLoS ONE

Article Title: The Phosphoinositide 3-Kinase p110α Isoform Regulates Leukemia Inhibitory Factor Receptor Expression via c-Myc and miR-125b to Promote Cell Proliferation in Medulloblastoma

doi: 10.1371/journal.pone.0123958

Figure Lengend Snippet: (a) Tumors formed on (CAM) were treated with 20μg/ml of rh-LIFα recombinant or phosphate-buffered saline for 4 consecutive days. Quantification of tumor volume changes before and after treatment is shown. Lines indicate the mean of each group, *P<0.05 compared with control treatment. (b) The expression of LIFRα downstream targets was analyzed upon treatment for 10, 30, 240 min with 20 μg/ml of rh-LIFRα. β-actin was used as a loading control.

Article Snippet: The Anti-human LIFRα antibody (AF-249-NA) and the recombinant rh-LIF Rα (7487-LR) were purchased from R&D Systems (Minneapolis, MN, USA) and were diluted directly into the medium immediately before use.

Techniques: Recombinant, Saline, Control, Expressing