rabbit anti mouse lif receptor lifr antibody  (Santa Cruz Biotechnology)

Journal: Cancers

Article Title: Gp130-Mediated STAT3 Activation Contributes to the Aggressiveness of Pancreatic Cancer through H19 Long Non-Coding RNA Expression

doi: 10.3390/cancers14092055

Figure Lengend Snippet: Active gp130/STAT3 pathway in PANC-1 sphere cells. ( A ) Real-time qPCR analysis of IL-6 , LIF , IL-6R , LIFR , and gp130 in PANC-1 cells cultured in 2D (adherent) or 3D (sphere) conditions. The results presented are normalized to values obtained for adherent cells (value = 1). Results are presented as means ± SD from three independent experiments. ( B ) FACS analysis of IL-6R, LIFR, and gp130 expression in PANC-1 cells cultured in 2D (adherent) or 3D (sphere) conditions. Controls are indicated by thin lines with gray color. ( C ) Cell surface levels of IL-6R, LIFR, and gp130 expression in PANC-1 cells cultured in 2D (adherent) or 3D (sphere) conditions. Mean fluorescence intensities (MFIs) relative to those of adherent cells are presented. Results are presented as means ± SD from three independent experiments. ( D ) Western blot analysis of p-gp130, gp130, p-STAT3, and STAT3 was performed in PANC-1 cells cultured in 2D (adherent) or 3D (sphere) conditions. Relative band intensity is provided. ** p < 0.01.

Article Snippet: Cells were harvested, and dissociated single cells were incubated on ice for 30 min with PE-conjugated anti-human IL-6 receptor α (IL-6Rα) antibody (BioLegend, San Diego, CA), PE-conjugated anti-human LIF receptor (LIFR) antibody (Becton Dickinson, Franklin Lakes, NJ, USA), PE-conjugated anti-human gp130 antibody (BioLegend), or PE-conjugated isotype control (Becton Dickinson) diluted in FACS buffer (0.5% ( w / v ) BSA and 0.1% ( w / v ) sodium azide in PBS).

Techniques: Cell Culture, Expressing, Fluorescence, Western Blot

Journal: Cancers

Article Title: Gp130-Mediated STAT3 Activation Contributes to the Aggressiveness of Pancreatic Cancer through H19 Long Non-Coding RNA Expression

doi: 10.3390/cancers14092055

Figure Lengend Snippet: The correlation of gp130/STAT3 pathway-related factor expression with overall survival and H19 expression correlated with gp130/STAT3 pathway-related factors in patients with PDAC. Each survival curve was presented according to the online database GEPIA. ( A ) IL-6 , ( B ) LIF , ( C ) IL-6R , ( D ) LIFR , ( E ) gp130 (IL-6ST) , ( F ) JAK1 , ( G ) STAT3 , ( H ) TGF β-RII (TGFBR2) , ( I ) MT1-MMP (MMP14) , and ( J ) H19 . ( K ) The correlation of H19 and gp130/STAT3 pathway-related factor expression in patients with PDAC was assessed using Spearman rank correlation analysis according to the online database GEPIA.

Article Snippet: Cells were harvested, and dissociated single cells were incubated on ice for 30 min with PE-conjugated anti-human IL-6 receptor α (IL-6Rα) antibody (BioLegend, San Diego, CA), PE-conjugated anti-human LIF receptor (LIFR) antibody (Becton Dickinson, Franklin Lakes, NJ, USA), PE-conjugated anti-human gp130 antibody (BioLegend), or PE-conjugated isotype control (Becton Dickinson) diluted in FACS buffer (0.5% ( w / v ) BSA and 0.1% ( w / v ) sodium azide in PBS).

Techniques: Expressing

Journal: Cancers

Article Title: Gp130-Mediated STAT3 Activation Contributes to the Aggressiveness of Pancreatic Cancer through H19 Long Non-Coding RNA Expression

doi: 10.3390/cancers14092055

Figure Lengend Snippet: Schematic representation of autocrine/paracrine IL-6 or the LIF/gp130/STAT3 pathway in PDAC sphere cells. In PDAC sphere cells in which CSCs are enriched (CSC-like cells), binding of autocrine/paracrine IL-6 or LIF to each receptor (IL-6R or LIFR, respectively) induces gp130 homodimerization or LIFR/gp130 complex formation to thereby activating JAKs followed by phosphorylation of gp130, ultimately leading to STAT3 activation. This pathway contributes to the maintenance of stemness features and expression of MT1-MMP and TGFβ-RII. Expression of TGFβ-RII via the gp130/STAT3 pathway affects TGF-β1/Smad signaling to promote EMT induction. Additionally, p-STAT3 can access the active promoter region of H19 and contribute to its transcription. Therefore, autocrine/paracrine IL-6 or the LIF/gp130/STAT3 pathway in PDAC CSC-like cells is believed to eventually lead to invasion and metastasis, both of which are hallmarks of malignancy.

Article Snippet: Cells were harvested, and dissociated single cells were incubated on ice for 30 min with PE-conjugated anti-human IL-6 receptor α (IL-6Rα) antibody (BioLegend, San Diego, CA), PE-conjugated anti-human LIF receptor (LIFR) antibody (Becton Dickinson, Franklin Lakes, NJ, USA), PE-conjugated anti-human gp130 antibody (BioLegend), or PE-conjugated isotype control (Becton Dickinson) diluted in FACS buffer (0.5% ( w / v ) BSA and 0.1% ( w / v ) sodium azide in PBS).

Techniques: Binding Assay, Phospho-proteomics, Activation Assay, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Leukemia Inhibitory Factor Induces Proopiomelanocortin via CRH/CRHR Pathway in Mouse Trophoblast

doi: 10.3389/fcell.2021.618947

Figure Lengend Snippet: CRH and LIF expression in mouse placenta during development. (A) Representative immunoblots and (B) quantification of CRH and LIF protein levels in mouse placenta at 13.5, 15.5, and 17.5 dpc. The housekeeping protein β-actin (42 kDa) was used as a loading control. Densitometry reveals a significant increase in CRH peptide and LIF at day 13.5 dpc in mouse placenta ( n = 4 of each group), with each point standardized to 17.5 dpc. All data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01 vs. control (Student’s t -test). (C) Placental sections obtained from mice at 13.5, 15.5, and 17.5 dpc underwent immunofluorescence staining using a CRH-specific antibody ( left panel ) and LIFR-specific antibody ( right panel ). Nuclei were stained with Hoechst 33342 ( blue ). Scale bar 50 μm. Dec decidua, JC junctional zone, Lab labyrinth, Cont negative control at13.5 dpc.

Article Snippet: Sections were incubated overnight at 4°C with a rabbit anti-mouse CRF antibody (2 mg/mL; Bioss; RRID:AB_10885736 ) and a rabbit anti-mouse LIF receptor (LIFR) antibody (0.2 mg/mL; Santa Cruz Biotechnology) after washing with PBS.

Techniques: Expressing, Western Blot, Control, Immunofluorescence, Staining, Negative Control

Journal: Journal of Neuroinflammation

Article Title: Inhibition of hypothalamic leukemia inhibitory factor exacerbates diet-induced obesity phenotype

doi: 10.1186/s12974-017-0956-9

Figure Lengend Snippet: The anatomical and cellular distribution of LIF and LIF receptor in the hypothalamus of mice. Hypothalamic frozen sections (5.0 μm thick) were prepared from 5-week-old mice fed on chow. Immunofluorescence staining was performed using antibodies against LIF ( a , c , e , g ), LIF receptor (LIFR) ( b , d , f , h ), NPY ( a , b ), POMC ( c , d ), GFAP ( e , f ), and IBA1 ( g , h ). Nuclei was labeled using DAPI. In some panels, a high magnification image depicts details of cells (arrows). Color code and magnifications are presented in the panels. Figures are representative of three independent experiments

Article Snippet: Sections were rinsed with PBS and blocked in a solution containing 5% normal serum and 0.2% Tween in phosphate buffered saline for 1 h at room temperature followed by incubation at 4 °C overnight with antibodies against LIF (ab113262, rabbit polyclonal, 1:500, ABCAM, Cambridge, UK) or LIF receptor (sc659, rabbit polyclonal, 1:200, Santa Cruz Biotechnology, Inc.) with ionized calcium binding adaptor molecule 1 (IBA-1, sc28530, goat polyclonal, 1:200, Santa Cruz Biotechnology, Inc.) or neuropeptide Y (NPY, sc133080, mouse monoclonal, 1:200, Santa Cruz Biotechnology, Inc.) or POMC (ab32893, goat polyclonal, 1:500, Abcam, Cambridge, UK) or glial fibrillary acidic protein (GFAP, ab4648, mouse monoclonal, 1:500, Abcam, Cambridge, UK) or alpha-MSH (AB5087, sheep polyclonal, 1:500, Millipore, Billerica, Massachusetts, USA) in a blocking buffer (1% bovine serum albumin in PBS-Tween).

Techniques: Immunofluorescence, Staining, Labeling

Journal: Journal of Neuroinflammation

Article Title: Inhibition of hypothalamic leukemia inhibitory factor exacerbates diet-induced obesity phenotype

doi: 10.1186/s12974-017-0956-9

Figure Lengend Snippet: Impact of inhibiting hypothalamic LIF on inflammatory markers. a Schematic representation of the protocol employed to immunoneutralize hypothalamic LIF. b The expression of LIF protein was evaluated by immunoblot in the hypothalamus of obesity-resistant (OR) mice treated with non-immune IgG or anti-LIF antibody for 15 days. Real-time PCR was employed to determine the relative expression of transcripts encoding for TNF-α ( c ), IL-1β ( d ), CX3CL1 (fractalkine) ( e ), cxcl1 ( f ) IL-6 ( g ), and IL-10 ( h ) ccl20 ( i ) in the hypothalamus of obesity-prone (OP) mice treated with non-immune IgG or OR mice treated either with non-immune IgG or anti-LIF antibody. In b – g , n = 5; * p < 0.05 vs. OP IgG and § p < 0.05 vs. OR IgG

Article Snippet: Sections were rinsed with PBS and blocked in a solution containing 5% normal serum and 0.2% Tween in phosphate buffered saline for 1 h at room temperature followed by incubation at 4 °C overnight with antibodies against LIF (ab113262, rabbit polyclonal, 1:500, ABCAM, Cambridge, UK) or LIF receptor (sc659, rabbit polyclonal, 1:200, Santa Cruz Biotechnology, Inc.) with ionized calcium binding adaptor molecule 1 (IBA-1, sc28530, goat polyclonal, 1:200, Santa Cruz Biotechnology, Inc.) or neuropeptide Y (NPY, sc133080, mouse monoclonal, 1:200, Santa Cruz Biotechnology, Inc.) or POMC (ab32893, goat polyclonal, 1:500, Abcam, Cambridge, UK) or glial fibrillary acidic protein (GFAP, ab4648, mouse monoclonal, 1:500, Abcam, Cambridge, UK) or alpha-MSH (AB5087, sheep polyclonal, 1:500, Millipore, Billerica, Massachusetts, USA) in a blocking buffer (1% bovine serum albumin in PBS-Tween).

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction

Journal: Journal of Neuroinflammation

Article Title: Inhibition of hypothalamic leukemia inhibitory factor exacerbates diet-induced obesity phenotype

doi: 10.1186/s12974-017-0956-9

Figure Lengend Snippet: Body mass, body composition, and caloric intake in mice treated with anti-LIF antibody. Mice were treated according to the protocol presented in Fig. a, and parameters were determined throughout the experimental period. a Body mass and b body mass variation. c Relative fat mass and d relative fat-free mass, as determined by body densitometry at the end of the experimental period. e Daily energy intake and f cumulative energy intake during the experimental period. In all experiments n = 5; * p < 0.05 vs. OP IgG and § p < 0.05 vs. OR IgG. OP IgG, obesity-prone mice treated with non-immune IgG; OR IgG, obesity-resistant mice treated with non-immune IgG; OR anti-LIF, obesity-resistant mice treated with anti-LIF antibody

Article Snippet: Sections were rinsed with PBS and blocked in a solution containing 5% normal serum and 0.2% Tween in phosphate buffered saline for 1 h at room temperature followed by incubation at 4 °C overnight with antibodies against LIF (ab113262, rabbit polyclonal, 1:500, ABCAM, Cambridge, UK) or LIF receptor (sc659, rabbit polyclonal, 1:200, Santa Cruz Biotechnology, Inc.) with ionized calcium binding adaptor molecule 1 (IBA-1, sc28530, goat polyclonal, 1:200, Santa Cruz Biotechnology, Inc.) or neuropeptide Y (NPY, sc133080, mouse monoclonal, 1:200, Santa Cruz Biotechnology, Inc.) or POMC (ab32893, goat polyclonal, 1:500, Abcam, Cambridge, UK) or glial fibrillary acidic protein (GFAP, ab4648, mouse monoclonal, 1:500, Abcam, Cambridge, UK) or alpha-MSH (AB5087, sheep polyclonal, 1:500, Millipore, Billerica, Massachusetts, USA) in a blocking buffer (1% bovine serum albumin in PBS-Tween).

Techniques:

Journal: Journal of Neuroinflammation

Article Title: Inhibition of hypothalamic leukemia inhibitory factor exacerbates diet-induced obesity phenotype

doi: 10.1186/s12974-017-0956-9

Figure Lengend Snippet: Respirometry and spontaneous activity in mice treated with anti-LIF antibody. Mice were treated according to the protocol presented in Fig. a and parameters were determined at the end of the experimental period. In respirometry, vO 2 ( a , b ) and vCO 2 ( c , d ) were measured during a 24-h period ( a , c ) and the means during light and dark periods were calculated ( b , d ). The respiratory quotient (RQ) was calculated during the 24-h period ( e ) and means were obtained during the light and dark periods ( f ). Spontaneous activity was determined during a 24-h period ( g ). In all experiments n = 5; * p < 0.05. OP IgG, obesity-prone mice treated with non-immune IgG; OR IgG, obesity-resistant mice treated with non-immune IgG; OR anti-LIF, obesity-resistant mice treated with anti-LIF antibody

Article Snippet: Sections were rinsed with PBS and blocked in a solution containing 5% normal serum and 0.2% Tween in phosphate buffered saline for 1 h at room temperature followed by incubation at 4 °C overnight with antibodies against LIF (ab113262, rabbit polyclonal, 1:500, ABCAM, Cambridge, UK) or LIF receptor (sc659, rabbit polyclonal, 1:200, Santa Cruz Biotechnology, Inc.) with ionized calcium binding adaptor molecule 1 (IBA-1, sc28530, goat polyclonal, 1:200, Santa Cruz Biotechnology, Inc.) or neuropeptide Y (NPY, sc133080, mouse monoclonal, 1:200, Santa Cruz Biotechnology, Inc.) or POMC (ab32893, goat polyclonal, 1:500, Abcam, Cambridge, UK) or glial fibrillary acidic protein (GFAP, ab4648, mouse monoclonal, 1:500, Abcam, Cambridge, UK) or alpha-MSH (AB5087, sheep polyclonal, 1:500, Millipore, Billerica, Massachusetts, USA) in a blocking buffer (1% bovine serum albumin in PBS-Tween).

Techniques: Activity Assay

Journal: Journal of Neuroinflammation

Article Title: Inhibition of hypothalamic leukemia inhibitory factor exacerbates diet-induced obesity phenotype

doi: 10.1186/s12974-017-0956-9

Figure Lengend Snippet: Determination of glucose tolerance in mice treated with anti-LIF antibody. Mice were treated according to the protocol presented in Fig. a and parameters were determined at the end of the experimental period. In the glucose tolerance test (GTT), blood glucose variation was measured from time 0 to 120 min ( a ) and the area under the glucose curve (AUC) was calculated ( b ). In insulin-tolerance test (ITT) blood glucose variation was measured form time 0 to 30 min ( c ) and the constant for glucose decay (kITT) was calculated ( d ). In all experiments n = 5; * p < 0.05 vs. OP IgG and § p < 0.05 vs. OR IgG. In b , p = 0.052 vs. OP IgG; in d , p = 0.075 vs. OR IgG. OP IgG, obesity-prone mice treated with non-immune IgG; OR IgG, obesity-resistant mice treated with non-immune IgG; OR anti-LIF, obesity-resistant mice treated with anti-LIF antibody

Article Snippet: Sections were rinsed with PBS and blocked in a solution containing 5% normal serum and 0.2% Tween in phosphate buffered saline for 1 h at room temperature followed by incubation at 4 °C overnight with antibodies against LIF (ab113262, rabbit polyclonal, 1:500, ABCAM, Cambridge, UK) or LIF receptor (sc659, rabbit polyclonal, 1:200, Santa Cruz Biotechnology, Inc.) with ionized calcium binding adaptor molecule 1 (IBA-1, sc28530, goat polyclonal, 1:200, Santa Cruz Biotechnology, Inc.) or neuropeptide Y (NPY, sc133080, mouse monoclonal, 1:200, Santa Cruz Biotechnology, Inc.) or POMC (ab32893, goat polyclonal, 1:500, Abcam, Cambridge, UK) or glial fibrillary acidic protein (GFAP, ab4648, mouse monoclonal, 1:500, Abcam, Cambridge, UK) or alpha-MSH (AB5087, sheep polyclonal, 1:500, Millipore, Billerica, Massachusetts, USA) in a blocking buffer (1% bovine serum albumin in PBS-Tween).

Techniques:

Journal: Journal of Neuroinflammation

Article Title: Inhibition of hypothalamic leukemia inhibitory factor exacerbates diet-induced obesity phenotype

doi: 10.1186/s12974-017-0956-9

Figure Lengend Snippet: The impact of pair feeding on the phenotype of obesity-resistant mice treated with anti-LIF antibody . Mice were treated with a protocol similar to the one presented in Fig. a, except that a group was submitted to pair feeding with the obesity-resistant mice treated with IgG throughout the experimental period. At the end of the experimental period, body mass variation was determined ( a ). In the glucose tolerance test (GTT), blood glucose variation was measured from time 0 to 120 min ( b ) and the area under the glucose curve (AUC) was calculated ( c ). In insulin-tolerance test (ITT), blood glucose variation was measured form time 0 to 30 min ( d ) and the constant for glucose decay (kITT) was calculated ( e ). In all experiments n = 5. In a , p = 0.053 vs. OR IgG; * p < 0.05 vs. OR anti-LIF AD. In c , * p < 0.05 vs. OR anti-LIF AD. In e , * p < 0.05 vs. OR IgG; p = 0.056 vs. OR anti-LIF AD

Article Snippet: Sections were rinsed with PBS and blocked in a solution containing 5% normal serum and 0.2% Tween in phosphate buffered saline for 1 h at room temperature followed by incubation at 4 °C overnight with antibodies against LIF (ab113262, rabbit polyclonal, 1:500, ABCAM, Cambridge, UK) or LIF receptor (sc659, rabbit polyclonal, 1:200, Santa Cruz Biotechnology, Inc.) with ionized calcium binding adaptor molecule 1 (IBA-1, sc28530, goat polyclonal, 1:200, Santa Cruz Biotechnology, Inc.) or neuropeptide Y (NPY, sc133080, mouse monoclonal, 1:200, Santa Cruz Biotechnology, Inc.) or POMC (ab32893, goat polyclonal, 1:500, Abcam, Cambridge, UK) or glial fibrillary acidic protein (GFAP, ab4648, mouse monoclonal, 1:500, Abcam, Cambridge, UK) or alpha-MSH (AB5087, sheep polyclonal, 1:500, Millipore, Billerica, Massachusetts, USA) in a blocking buffer (1% bovine serum albumin in PBS-Tween).

Techniques:

Journal: PLoS ONE

Article Title: The Inhibition of N-Glycosylation of Glycoprotein 130 Molecule Abolishes STAT3 Activation by IL-6 Family Cytokines in Cultured Cardiac Myocytes

doi: 10.1371/journal.pone.0111097

Figure Lengend Snippet: Neonatal rat cardiomyocytes were treated with the indicated concentrations of Tm for 8 hrs (A), or with 2 µg/mL of Tm for the indicated times (B), respectively. After each treatment, cell lysates were prepared and immunoblotted with anti-gp130, LIFR or IL-11Rα antibody, respectively. More than three independent experiments were performed with similar results and representative images were shown.

Article Snippet: Anti-STAT3 (sc-7179), anti-gp130 (sc-656), anti-phospho-JAK1 (sc-16773), anti-JAK1 (sc-7228), anti-LIF receptor (LIFR) (sc-659), anti-IL-11 receptor α (IL-11Rα) (sc-993) and anti-PTP1B (sc-1718) antibodies were obtained from Santa Cruz Biotechnology.

Techniques: