Journal: Endocrinology

Article Title: Leukemia Inhibitory Factor Is Necessary for Ovulation in Female Rhesus Macaques

doi: 10.1210/en.2016-1283

Figure Lengend Snippet: Antibody Table

Article Snippet: Any white balance adjustment was made uniformly across all images. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Protein Target Antigen Sequence Antibody Name Manufacturer and Catalog Number Species Raised in; Clonality Dilution Used LIFR KLH conjugated from the middle of human LIFR LIFR/CD118 Bioss, bs-1458R Rabbit; polyclonal 1:200 pY705-STAT3 Phosphopeptide containing human STAT3 Y705 site Human phosho-STAT3 (Y705) R&D Systems, AF4607 Rabbit; polyclonal 1:50 IL6ST Phosphorylation site of serine 782 CD130 (gp130) Abcam, ab59389 Rabbit; polyclonal 1:100 JAK1 Phosphorylation site of tyrosine 1022 JAK1 Abcam, ab47435 Rabbit; polyclonal 1:100 STAT3 Residues surrounding serine 727 of human STAT3 STAT3 Abcam, ab32500 Rabbit; monoclonal 1:100 pS727-STAT3 Human STAT3 700 to the C terminus STAT3 (phospho-S727) Abcam, ab32143 Rabbit; monoclonal 1:650 Isotype control Proprietary isotype control Rabbit IgG Abcam, ab172730 Rabbit; monoclonal 1:50, 100, 200, and 650 Open in a separate window Antibody Table Laparoscopic and histological evaluation of follicle rupture The presence or absence of an ovulatory stigmata was determined by laparoscopy at 72 hours after intrafollicular injection of vehicle (n = 3) or sLIFR (n = 4) and hCG ( 16 ).

Techniques: Sequencing, Control

Journal: Frontiers in Microbiology

Article Title: Protective Vaccination against Blood-Stage Malaria of Plasmodium chabaudi : Differential Gene Expression in the Liver of Balb/c Mice toward the End of Crisis Phase

doi: 10.3389/fmicb.2016.01087

Figure Lengend Snippet: Genes down-regulated more than 10-fold ( p < 0.01) in the liver of vaccinated mice infected with P. chabaudi on day 11 p.i. (Vd11) in comparison to constitutive expression on day 0 p.i. (Vd0) .

Article Snippet: Using High Capacity cDNA Reverse Transcription Kit (Life Technologies) and TaqMan mRNA assays (Life Technologies) we performed reverse transcription of mRNAs coding for the following proteins: RHD (assay ID: Mm00456910_m1), ERMAP (Mm_00469273_m1), ADD2 (Mm00478923_m1), ANK1 (Mm00482889_m1), PLK1 (Mm00440924_g1), CCNA2 (Mm00438063_m1), LIFR (Mm00442942_m1), and CD163 (Mm_00474091_m1).

Techniques: Infection, Comparison, Expressing, Bacteria, Binding Assay, Activity Assay, Blocking Assay, Translocation Assay, Clinical Proteomics, Membrane

Journal: Journal of neuropathology and experimental neurology

Article Title: Effects of oncostatin M on human cerebral endothelial cells and expression in inflammatory brain lesions.

doi: 10.1093/jnen/60.11.1087

Figure Lengend Snippet: Fig. 2. HCEC transcribe mRNA for gp130, OSMR-b, LIFR, and IL-6R, but not for CNTFR. RT-PCR was performed on RNA extracted from HCEC and reverse-transcribed (RT1) using oligo(dT) primers. RNA samples not submitted to reverse transcription were used as negative controls (RT2). PCR results for gp130, LIFR, and OSMR-b are shown in the upper panel. Note that the expression level of LIFR was below that of gp130 and OSMR-b even when a 3-fold (33) higher amount of cDNA was used as starting material in the RT-PCR. PCR amplicons of CNTFR, IL-6R and b-actin (b-A), which was used as an internal control, are shown in the lower panel. RNA extracted from early postmortem human brain (hu Brain) served as a positive control for CNTFR. Identical results were obtained in all 5 HCEC preparations analyzed. Length of bands from DNA size marker (M, 1 kb DNA ladder, Gibco BRL) is indicated on the left (A). Surface detection of gp130 and OSMR-b, but not of LIFR, on HCEC. FACS analysis was performed as described in Materials and Methods to determine surface expression of gp130, OSMR-b, and LIFR on HCEC. Histograms show stain- ing with the respective receptor antibody (shaded) underlaid with the isotype-matched control (single line). Hela cells were

Article Snippet: For FACS analysis of adhesion molecules and OSM receptor subunits, HCEC were detached with 0.1% Trypsin/EDTA (Sigma), washed with PBS containing 0.1% BSA (Sigma) and 0.1% D ow nloaded from https://academ ic.oup.com /jnen/article/60/11/1087/2916229 by guest on 25 August 2024 J Neuropathol Exp Neurol, Vol 60, November, 2001 sodium azide, and incubated with primary mouse monoclonal antibodies against ICAM-1 (1:30; Bender MedSystems, Vienna, Austria), VCAM-1 (1:30; Biosource), gp130 (1:10), OSMRb (1:10, Santa Cruz Biotechnology, Santa Cruz, CA), and LIFR (1:10, Santa Cruz Biotechnology) for 30 min at 48C.

Techniques: Reverse Transcription Polymerase Chain Reaction, Reverse Transcription, Expressing, Control, Positive Control, Marker, Staining

Journal: Nature Communications

Article Title: A targetable LIFR−NF-κB−LCN2 axis controls liver tumorigenesis and vulnerability to ferroptosis

doi: 10.1038/s41467-021-27452-9

Figure Lengend Snippet: a Correlation between LIFR expression and erastin sensitivity, based on the liver cancer cell lines ( n = 22) from CTRP. Dose responses are normalized area under curve values. The linear relationship was determined by a two-tailed Pearson correlation analysis. b , c Lifr-knockout ( b ) and LIFR-overexpressing ( c ) PHM cells were treated with DMSO (vehicle), cystine starvation, erastin (10 μM), or RSL3 (0.1 μM) for 24 h. Cell death was measured by PI staining. n = 3 wells. d Kaplan−Meier curves of overall survival of Lifr fl/fl and Lifr fl/fl ;Alb-Cre mice that received sorafenib treatment 7 days after hydrodynamic injection of plasmids expressing the Sleeping Beauty transposase, myrAKT-IRES-luciferase, and RasV12. Sorafenib was administered 6 days a week. Statistical significance was determined by a log-rank test. n = 6 mice. e Photon flux of Lifr fl/fl and Lifr fl/fl ;Alb-Cre mice that received sorafenib treatment 7 days after hydrodynamic injection of plasmids expressing the Sleeping Beauty transposase, myrAKT-IRES-luciferase, and RasV12. Sorafenib was administered 6 days a week. Statistical significance was determined by a two-tailed unpaired t -test. n = 6 mice. f , g Immunohistochemical staining ( f ) and quantification ( g ) of 4-HNE in livers of the mice described in e . n = 6 mice. Scale bars, 200 μm. h , i Liver weight ( h ) and liver-to-body weight ratio ( i ) of C57BL/6 mice that received control adenovirus or LIFR-expressing adenovirus 3 days and 17 days after hydrodynamic injection of plasmids expressing the Sleeping Beauty transposase, myrAKT, and RasV12. One week after plasmid injection, mice received 30 mg kg −1 sorafenib and/or 10 mg kg −1 liproxstatin-1, 6 days a week for 4 weeks. n = 4, 4, 3, 4, 5, 5, 5, and 7 mice from left to right. j , k Immunohistochemical staining ( j ) and quantification ( k ) of 4-HNE in livers of the mice described in h . n = 4, 4, 4, 4, 4, 5, 5, and 7 mice from left to right. Scale bars, 200 μm. Statistical significance in b , c , g – i , and k was determined by a two-tailed unpaired t -test. Error bars are s.e.m. Source data are provided as a Source Data file.

Article Snippet: CRISPR-Cas9-mediated Lifr-knockout constructs were purchased from Santa Cruz Biotechnology (sc-421433).

Techniques: Expressing, Two Tailed Test, Knock-Out, Staining, Injection, Luciferase, Immunohistochemical staining, Plasmid Preparation

Journal: Nature Communications

Article Title: A targetable LIFR−NF-κB−LCN2 axis controls liver tumorigenesis and vulnerability to ferroptosis

doi: 10.1038/s41467-021-27452-9

Figure Lengend Snippet: a Volcano plot of genes upregulated (red) or downregulated (blue) in Lifr fl/fl ;Alb-Cre mice ( n = 3) relative to Lifr fl/fl mice ( n = 2). Statistical analysis of RNA-seq data was performed using Cuffdiff and P values are false discovery rate (FDR)-adjusted. b Cytokine arrays of the conditioned medium of Lifr-knockout PHM cells. Boxed: the top five upregulated (red) and downregulated (blue) cytokines. c qPCR of Lcn2 in Lifr-knockout PHM cells. n = 3 samples. d ELISA of lipocalin 2 in the conditioned medium of Lifr-knockout PHM cells. n = 5, 4, and 5 wells from left to right. e ELISA of lipocalin 2 in the serum of 3-month-old Lifr fl/fl and Lifr fl/fl ;Alb-Cre mice. n = 4 mice. f Pathway analysis of Lifr-knockout PHM cells with or without LIFR add-back. g , h Immunoblotting of p-p65, p65, and LIFR in LIFR-knockdown ( g ) and LIFR-overexpressing ( h ) Mahlavu cells. i , j Pathway analysis of livers of Lifr fl/fl and Lifr fl/fl ;Alb-Cre mice that received hydrodynamic injection of plasmids expressing the Sleeping Beauty transposase and oncogenes ( i : β-catenin + YAP; j : myrAKT + RasV12). k , l qPCR of Lcn2 in livers described in i and j , respectively. n = 3 samples per mouse; n = 2 mice per group. m , n Immunohistochemical staining ( m ) and quantification ( n ) of Lcn2 in livers described in i and j , respectively. n = 6 mice. o Immunoblotting of Lifr, p-p65, p65, p-Stat3, Stat3, and Gapdh in livers of Lifr fl/fl and Lifr fl/fl ;Cre-ERT2 mice, 28 days after hydrodynamic injection of plasmids expressing the Sleeping Beauty transposase, myrAKT, and RasV12. From day 7, all mice received 5-day tamoxifen treatment. p , q Immunohistochemical staining ( p ) and quantification ( q ) of Lcn2 in livers of the mice that received control or LIFR-expressing adenovirus 3 days and 17 days after hydrodynamic injection of plasmids expressing the Sleeping Beauty transposase, myrAKT, and RasV12. Scale bars, 200 μm. n = 4 mice. Statistical significance in c – e , n , and q was determined by a two-tailed unpaired t -test. Error bars are s.e.m. Source data are provided as a Source Data file.

Article Snippet: CRISPR-Cas9-mediated Lifr-knockout constructs were purchased from Santa Cruz Biotechnology (sc-421433).

Techniques: RNA Sequencing Assay, Knock-Out, Enzyme-linked Immunosorbent Assay, Western Blot, Injection, Expressing, Immunohistochemical staining, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: A targetable LIFR−NF-κB−LCN2 axis controls liver tumorigenesis and vulnerability to ferroptosis

doi: 10.1038/s41467-021-27452-9

Figure Lengend Snippet: a HEK293T cells were transfected with HA-FLAG-SHP1 and SFB-tagged GFP or LIFR. LIFR-SFB protein was pulled down with S-protein beads, followed by immunoblotting with antibodies against FLAG and HA. b HEK293T cells were transfected with MYC-SHP2 and SFB-tagged GFP or LIFR. LIFR-SFB protein was pulled down with S-protein beads, followed by immunoblotting with antibodies against FLAG and MYC. c HEK293T SFB-GFP and SFB-LIFR stable cell lines were infected with the scrambled (Scr) or sh-SHP1 lentivirus, followed by transfection with a K63-specific mutant of His-Xpress-ubiquitin (Ub). 48 h later, cells were subjected to pulldown with nickel beads and immunoblotting with antibodies against TRAF6 and Xpress. d Control and LIFR-overexpressing PLC/PRF/5 cells were transduced with SHP1 shRNA and immunoblotted with the indicated antibodies. e Control (Scr) and LIFR-knockdown HEK293T cells were transfected with FLAG-TRAF6. 48 h later, cells were immunoprecipitated with a FLAG-specific antibody and immunoblotted with antibodies against LIFR, SHP1, and FLAG. f , g qPCR of LCN2 , LIFR , and RELA in HEK293T ( f ) and PLC/PRF/5 ( g ) cells transduced with LIFR shRNA alone or in combination with p65 shRNA. n = 3 technical replicates. h qPCR of Lcn2 , Lifr , and RelA in control and Lifr-knockout PHM cells transduced with the scrambled shRNA (Scr) or p65 shRNA. n = 3 technical replicates. i , j Immunohistochemical staining ( i ) and quantification ( j ) of Lcn2 in livers from Lifr fl/fl (F/F) and Lifr fl/fl ;Alb-Cre (LKO) mice, 57 days after hydrodynamic injection of plasmids expressing the Sleeping Beauty transposase, myrAKT, RasV12, and shRNA (sh-p65, sh-Lcn2, or scrambled). n = 10, 8, 10, and 12 mice from left to right. Scale bars, 200 μm. k , l Liver weight ( k ) and liver-to-body weight ratio ( l ) of the mice described in i and j . n = 10, 8, 10, and 12 mice from left to right. m H&E staining of livers described in i and j . Scale bars, 300 μm. Statistical significance in f − h and j − l was determined by a two-tailed unpaired t -test. Error bars are s.e.m. Source data are provided as a Source Data file.

Article Snippet: CRISPR-Cas9-mediated Lifr-knockout constructs were purchased from Santa Cruz Biotechnology (sc-421433).

Techniques: Transfection, Western Blot, Stable Transfection, Infection, Mutagenesis, Transduction, shRNA, Immunoprecipitation, Knock-Out, Immunohistochemical staining, Staining, Injection, Expressing, Two Tailed Test

Journal: Nature Communications

Article Title: A targetable LIFR−NF-κB−LCN2 axis controls liver tumorigenesis and vulnerability to ferroptosis

doi: 10.1038/s41467-021-27452-9

Figure Lengend Snippet: a qPCR of Lifr and Lcn2 in Lifr-knockout PHM cells transduced with the scrambled (Scr) or Lcn2 shRNA. n = 3 samples. b Lifr-knockout PHM cells were transduced with Lcn2 shRNA and treated with 10 µM erastin or 20 µM sorafenib. Cell viability was determined by a CCK8 assay. n = 5 wells. c Fe 2+ levels in liver tissues of Lifr fl/fl and Lifr fl/fl ;Alb-Cre mice in the absence or presence of hydrodynamic injection of plasmids expressing the Sleeping Beauty transposase, myrAKT, and RasV12. n = 4 mice. d Fe 2+ levels in control and Lifr-knockout PHM cells transduced with the scrambled (Scr) or Lcn2 shRNA. n = 3 wells. e Malondialdehyde (MDA) levels in control and Lifr-knockout PHM cells transduced with the scrambled (Scr) or Lcn2 shRNA. n = 5 wells. f Glutathione (GSH) levels in control and Lifr-knockout PHM cells transduced with the scrambled (Scr) or Lcn2 shRNA. n = 3 wells. g Immunoblotting of Lifr, Slc7a11, Fsp1, Gpx4, and Gapdh in control and Lifr-knockout PHM cells transduced with the scrambled (Scr) or Lcn2 shRNA. h Immunoblotting of LCN2, LIFR, p-p65, p65, and GAPDH in tumors generated from four PDX lines of HCC. i ELISA of lipocalin 2 in the serum collected from NSG mice bearing PDX line #5. Mice were treated with anti-LCN2 and sorafenib, alone or in combination. n = 7 mice. j , k Growth curves of tumors in NSG mice bearing PDX line #4 ( j ) or #5 ( k ). When tumors grew to 50–150 mm 3 , mice were treated with 100 μg anti-LCN2 and 30 mg kg −1 sorafenib, alone or in combination. The treatments were given 6 days a week for 4 weeks. Statistical significance was determined by a two-way ANOVA. n = 6 mice in j and n = 7 mice in k . l Endpoint tumor images of the mice described in j and k . Statistical significance in b – f and i was determined by a two-tailed unpaired t -test. Error bars are s.e.m. Source data are provided as a Source Data file.

Article Snippet: CRISPR-Cas9-mediated Lifr-knockout constructs were purchased from Santa Cruz Biotechnology (sc-421433).

Techniques: Knock-Out, Transduction, shRNA, CCK-8 Assay, Injection, Expressing, Western Blot, Generated, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: BMC Immunology

Article Title: Antagonist anti-LIF antibody derived from naive human scFv phage library inhibited tumor growth in mice

doi: 10.1186/s12865-024-00636-w

Figure Lengend Snippet: Characterization of antibody 1G11. ( A ) SPR affinity analysis of 1G11 binding to human, cynomolgus and mouse LIF. ( B ) Competitive ELISA and SPR analysis of 1G11. 1G11 competes with LIFR for binding to LIF, while 1G11 does not compete with gp130 for binding to LIF. ( C ) Western-bolt analysis for the inhibitory effect of 1G11 on LIF-induced p-STAT3

Article Snippet: The other recombinant protein sources were as follows: human LIF (Z02681, Genscript, China), biotinylated human LIF (LIF-H82E2, Acrobiosystems, China), mouse LIF (250-02, PeproTech, USA), cynomolgus LIF (RP1074Y-100, Kingfisher Biotech, USA), human LIFR-his (10,628-H08H, Sino Biological, China), and human gp130-his (10,974-HCCH, Sino Biological, China).

Techniques: Binding Assay, Competitive ELISA, Western Blot

Journal: Stem Cell Research & Therapy

Article Title: Comparative analysis of gene transcripts for cell signaling receptors in bone marrow-derived hematopoietic stem/progenitor cell and mesenchymal stromal cell populations

doi: 10.1186/scrt323

Figure Lengend Snippet: Differential gene expression between bone marrow-derived hematopoietic stem/progenitor cells and bone marrow-derived mesenchymal stromal cells

Article Snippet: Lifr-leukemia inhibitory factor receptor , Mm00442940_m1 , 17.89 ± 0.64 , 16.76 ± 0.36 , -2.3 , 0.0001.

Techniques: Gene Expression, Derivative Assay, Virus, Binding Assay

Journal: Cell stem cell

Article Title: LIF signaling regulates outer radial glial to interneuron fate during human cortical development

doi: 10.1016/j.stem.2023.08.009

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The conjugated antibody against LIFR (biotechne, #FAB249A) was added directly to the suspended cells (100 μl per 1 ml FACS buffer) and incubated for 30 minutes.

Techniques: Recombinant, Multiplex Assay, Software