Journal: Cell Death & Disease

Article Title: Leukemia inhibitory factor drives glucose metabolic reprogramming to promote breast tumorigenesis

doi: 10.1038/s41419-022-04820-x

Figure Lengend Snippet: A Ectopic LIF expression (left panel) or treatment with rhLIF (right) enhanced lactate production of different breast cancer cell lines analyzed by measuring lactate levels in the medium using a lactate assay kit. B LIF neutralization antibody (LIF neu-ab) blocked the promoting effect of LIF on lactate production in MCF7, MDA-MB 231 and T47D cells. C Knockdown of endogenous LIF decreased lactate production in MDA-MB 231 cells. D , E Ectopic LIF expression enhanced glycolysis rates ( D ), whereas knockdown of endogenous LIF decreased glycolysis rates ( E ) in different breast cancer cell lines as calculated by ECAR measured by using a Seahorse analyzer. F The diagram of glycolysis. G – I The fold change of inter-metabolites in glycolysis resulted from ectopic LIF expression in MCF7 and MDA-MB 231 cells ( G ), rhLIF treatment in MCF7 cells ( H ) or LIF knockdown in MDA-MB 231 cells ( I ) as determined by LC/MS metabolomics analysis. J The fold change of inter-metabolites in glycolysis induced by ectopic LIF expression in MCF7 xenograft tumors determined by LC/MS metabolomics analysis. Data are presented as mean ± SD ( n = 3/group). * p < 0.05; ** p < 0.01; *** p < 0.001; unpaired Student’s t -test.

Article Snippet: LIF neutralization antibody (AF-250-NA) was purchased from R&D Systems.

Techniques: Expressing, Lactate Assay, Neutralization, Knockdown, Liquid Chromatography with Mass Spectroscopy

Journal: Cell Death & Disease

Article Title: Leukemia inhibitory factor drives glucose metabolic reprogramming to promote breast tumorigenesis

doi: 10.1038/s41419-022-04820-x

Figure Lengend Snippet: A Glut1 mRNA levels were detected by quantitative real-time PCR (qPCR) in MCF7, MDA-MB 231, and T47D cells with or without ectopic LIF expression. B , C Ectopic LIF expression ( B ) or rhLIF treatment (100 ng/ml for 12 h) ( C ) promoted endogenous Glut1 PM translocation in MCF7, MDA-MB 231, and T47D cells as determined by Western-blot assays. D Knockdown of LIF decreased endogenous Glut1 PM translocation in MDA-MB 231 cells. E Ectopic LIF expression increased the PM translocation of ectopically expressed Myc-Glut1 in MCF7, MDA-MB 231 and T47D cells as determined by Western-blot assays. F LIF neutralization antibody (LIF neu-ab) largely abolished exogenous Glut1 PM translocation promoted by LIF. G The rhLIF treatment promoted exogenous Glut1 PM translocation in cells. H Knockdown of LIF decreased Myc-Glut1 PM translocation in MDA-MB 231 cells. I Ectopic LIF expression promoted Myc-Glut1 PM translocation (left panels) while knockdown of LIF decreased Myc-Glut1 PM translocation (right panels) in MDA-MB 231 cells as determined by IF staining assays. Scale bar, 10 μm. J Ectopic LIF expression promoted the PM translocation of Myc-Glut1 in MCF7, MDA-MB 231, and T47D cells as determined by flow cytometry assays. Left panels: representative images of flow cytometry analysis. Right panels: quantifications of relative fluorescence intensity of Myc-Glut1 on the cell membrane normalized with total Myc-Glut1 fluorescence intensity in cells. In A , J data are presented as mean ± SD. n = 3/group. * p < 0.05; ** p < 0.01; NS: non-significant; unpaired Student’s t -test. Uncropped Wes t ern-blot images are shown in Supplementary Fig .

Article Snippet: LIF neutralization antibody (AF-250-NA) was purchased from R&D Systems.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Translocation Assay, Western Blot, Knockdown, Neutralization, Staining, Flow Cytometry, Fluorescence, Membrane

Journal: Comparative Hepatology

Article Title: Expression of leukemia inhibitory factor (LIF) and its receptor gp190 in human liver and in cultured human liver myofibroblasts. Cloning of new isoforms of LIF mRNA

doi: 10.1186/1476-5926-3-10

Figure Lengend Snippet: Detection of LIF transcripts in cultured human liver myofibroblasts. ( A ): Northern blot. Total RNA from cultured human liver myofibroblasts was hybridized with a cDNA probe to human LIF. A single 4.5 kb band was observed; ( B ) and ( C ): RT-PCR. Total RNA was subjected to reverse transcription then to PCR with the hLIF-D3/hLIF-N4 ( B ) or with the hLIF-M3/hLIF-N4 primers ( C ).

Article Snippet: A commercially available polyclonal antibody against human LIF (R&D Systems, Minneapolis, Minnesota, USA), and different monoclonal antibodies against gp190, previously described [ ], were used at concentrations optimised on control tissues.

Techniques: Cell Culture, Northern Blot, Reverse Transcription Polymerase Chain Reaction, Reverse Transcription

Journal: Comparative Hepatology

Article Title: Expression of leukemia inhibitory factor (LIF) and its receptor gp190 in human liver and in cultured human liver myofibroblasts. Cloning of new isoforms of LIF mRNA

doi: 10.1186/1476-5926-3-10

Figure Lengend Snippet: Sequence of LIF-D, M and T isoforms. Exons D, M and T are the 3 alternate first exons. Primers used for PCR are underlined. Primers hLIF-M2, M3 and M5 cover the same sequence but differ because of the presence or the absence of restriction sites.

Article Snippet: A commercially available polyclonal antibody against human LIF (R&D Systems, Minneapolis, Minnesota, USA), and different monoclonal antibodies against gp190, previously described [ ], were used at concentrations optimised on control tissues.

Techniques: Sequencing

Journal: Comparative Hepatology

Article Title: Expression of leukemia inhibitory factor (LIF) and its receptor gp190 in human liver and in cultured human liver myofibroblasts. Cloning of new isoforms of LIF mRNA

doi: 10.1186/1476-5926-3-10

Figure Lengend Snippet: RT-PCR analysis of LIF-M expression in various cell lines and in human liver. ( A ): LIF-M expression was analyzed with the hLIF-M2 and hLIF-3N primers: Line 1, human liver myofibroblasts; Line 2, HepG2; Line 3, Hep3B; Line 4, HuH7; Line 5, HEK293. Product sizes are shown in bp; ( B ): normal human liver samples. LIF-D expression was analyzed with the hLIF-D3 and hLIF-N4 primers in 4 different samples. The same samples also expressed LIF-M (not shown). Product sizes are shown in bp; ( C ): diseased human liver samples. In that case, LIF-M expression was analyzed with the hLIF-M3 and hLIF-4N primers in 4 cases of cirrhotic liver. The same samples also expressed LIF-D (not shown). Product sizes are shown in bp; ( D ): semi-quantitation of LIF-D and s-LIF-D expression in a human liver myofibroblasts sample. LIF-D expression was analyzed with the hLIF-D3 and hLIF-N4 primers. The left part shows the migration pattern of the PCR-amplified products with the number of cycles above and the size of the products indicated by arrows, in bp. The graph on the right shows the signal quantification. Similar results were obtained with LIF-M.

Article Snippet: A commercially available polyclonal antibody against human LIF (R&D Systems, Minneapolis, Minnesota, USA), and different monoclonal antibodies against gp190, previously described [ ], were used at concentrations optimised on control tissues.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Quantitation Assay, Migration, Amplification

Journal: PLoS ONE

Article Title: Leukemia Inhibitory Factor Enhances Endometrial Stromal Cell Decidualization in Humans and Mice

doi: 10.1371/journal.pone.0025288

Figure Lengend Snippet: A–D. LIF immunolocalized strongly to the decidua (d) and also to the luminal epithelium (le). E–H. LIFR immunolocalized strongly to the decidua. Bottom row, negative controls to sections above. The same negative is shown for both antibodies as the primary antibodies shown were both raised in goat. n = 3; m. myometrium; s, stroma.

Article Snippet: LIF and LIFR were immunolocalised as described above except that the non-immune block used was: LIF: 10% normal goat plus 2% normal mouse serum and LIFR: 10% normal horse and 2% mouse serum; and two LIF antibodies were used to confirm the specificity of LIF staining (rabbit anti-human LIF antibody as above and in and goat anti-human LIF antibody, R&D systems; 10 μg/ml).

Techniques:

Journal: Genes & Diseases

Article Title: Targeting LIF/LIFR signaling in cancer

doi: 10.1016/j.gendis.2021.04.003

Figure Lengend Snippet: Schematic depicting the drugs that inhibit LIF/LIFR downstream signaling. Anti LIF antibody MSC-1 or LIFR inhibitor EC359 can be used to directly interfere LIF/LIFR signaling. Targeting LIF/LIFR signaling using inhibitors of LIFR activated pathways including Jak1/STAT3 inhibitors, PI3K inhibitors or BRD inhibitors may also be useful to interfere LIF/LIFR signaling.

Article Snippet: Considering the importance of the LIF/LIFR pathway, Northern Biologics/Celgene recently developed a humanized Anti-LIF antibody (MSC-1) that blocks LIF signaling, and its utility is being tested in a phase I clinical trial to determine its safety and tolerability (ClinicalTrials.gov, NCT03490669 ).

Techniques: