Journal: Traffic (Copenhagen, Denmark)

Article Title: ENDOCYTOSIS OF LIGAND ACTIVATED SPHINGOSINE 1-PHOSPHATE RECEPTOR 1 MEDIATED BY THE CLATHRIN-PATHWAY

doi: 10.1111/tra.12343

Figure Lengend Snippet: (A-D) HeLa cells not expressing (A) or transiently expressing 3xHA-S1PR1-eGFP (B-D) were incubated with anti-HA antibody to label receptor at the cell surface and then follow their internalization upon incubation with DMSO only (Carrier) or the S1PR1 ligands (S1P or FTY720P). Fluorescent transferrin-Alexa Fluor 647 was used to monitor the clathrin-based receptor mediated endocytosis of transferrin. HA antibody was used to follow the uptake of S1P1R. Images for each condition correspond to a center plane obtained using spinning disc confocal microscopy from which quantification data was obtained. An acid wash step at the end of the experiment was used to remove the surface bound transferrin or HA antibodies. Scale bar 10 μm.

Article Snippet: Reagents Sphingosine-1-phosphate (S1P) and FTY720-phosphate (FTY720P) were from Echelon Bioscience (Salt Lake City, UT), poly-d-lysine (Sigma) and collagen (BD Biosciences).

Techniques: Expressing, Incubation, Confocal Microscopy

Journal: Traffic (Copenhagen, Denmark)

Article Title: ENDOCYTOSIS OF LIGAND ACTIVATED SPHINGOSINE 1-PHOSPHATE RECEPTOR 1 MEDIATED BY THE CLATHRIN-PATHWAY

doi: 10.1111/tra.12343

Figure Lengend Snippet: (A) Effect of clathrin or AP2 depletion in the uptake of S1PR1 in Hela cells transiently expressing 3xHA-S1P1-eGFP. Cells were depleted of clathrin or of its endocytic adaptor AP2 by transduction with lentiviruses encoding shRNA specific for clathrin heavy chain (CLC) or the μ2 subunit of AP2 (AP2); scrambled shRNA was used as a negative control. The cells were incubated with anti-HA antibody to follow the uptake of S1PR1 upon addition of DMSO only (Carrier) or the ligands (S1P or FTY720P). Transferrin-Alexa Fluor 647 was used to monitor its clathrin-based receptor mediated endocytosis. The representative images correspond to a center plane from which quantification data was obtained. An acid wash step at the end of the experiment was used to remove surface bound transferrin and anti-HA antibodies. Scale bar 10 μm.

Article Snippet: Reagents Sphingosine-1-phosphate (S1P) and FTY720-phosphate (FTY720P) were from Echelon Bioscience (Salt Lake City, UT), poly-d-lysine (Sigma) and collagen (BD Biosciences).

Techniques: Expressing, Transduction, shRNA, Negative Control, Incubation

Journal: Traffic (Copenhagen, Denmark)

Article Title: ENDOCYTOSIS OF LIGAND ACTIVATED SPHINGOSINE 1-PHOSPHATE RECEPTOR 1 MEDIATED BY THE CLATHRIN-PATHWAY

doi: 10.1111/tra.12343

Figure Lengend Snippet: (A) MEF cells from mice expressing (β-Arrestin1/2+/+) or not (β-Arrestin1/2−/−) β-arrestin1 and β-arrestin2 and stably expressing 3xHA-S1PR1-eGFP were incubated with anti-HA antibody in the presence of DMSO only (Carrier) or the ligands S1P or FTY720P. Images for each condition correspond to a center plane obtained using spinning disc confocal microscopy. An acid wash step at the end of the experiment was used to remove most of the surface bound anti-HA antibodies. Scale bar 10 μm.

Article Snippet: Reagents Sphingosine-1-phosphate (S1P) and FTY720-phosphate (FTY720P) were from Echelon Bioscience (Salt Lake City, UT), poly-d-lysine (Sigma) and collagen (BD Biosciences).

Techniques: Expressing, Stable Transfection, Incubation, Confocal Microscopy

Journal: Traffic (Copenhagen, Denmark)

Article Title: ENDOCYTOSIS OF LIGAND ACTIVATED SPHINGOSINE 1-PHOSPHATE RECEPTOR 1 MEDIATED BY THE CLATHRIN-PATHWAY

doi: 10.1111/tra.12343

Figure Lengend Snippet: HEK 293A cells stably expressing 3xHA-S1PR1-mCherry were incubated or not with anti-HA antibody to label receptor at the cell surface and then follow their internalization upon incubation with DMSO only (carrier) or the S1PR1 ligands (S1P or FTY720P). Fluorescent transferrin-Alexa Fluor 647 was used to monitor the clathrin-based receptor mediated endocytosis of transferrin. Data obtained using flow cytometry was from more than 10,000 cells analyzed per condition.

Article Snippet: Reagents Sphingosine-1-phosphate (S1P) and FTY720-phosphate (FTY720P) were from Echelon Bioscience (Salt Lake City, UT), poly-d-lysine (Sigma) and collagen (BD Biosciences).

Techniques: Stable Transfection, Expressing, Incubation, Flow Cytometry

Journal: Traffic (Copenhagen, Denmark)

Article Title: ENDOCYTOSIS OF LIGAND ACTIVATED SPHINGOSINE 1-PHOSPHATE RECEPTOR 1 MEDIATED BY THE CLATHRIN-PATHWAY

doi: 10.1111/tra.12343

Figure Lengend Snippet: (A) HeLa cells transiently expressing 3xHA-S1PR1-eGFP were incubated with anti-HA antibody in the absence (− dynasore-OH) or presence (+ dynasore-OH) of the modified more potent cell permeable inhibitor of dynamin, dynasore-OH. Following brief 15 min incubation with dynasore-OH, the media was replaced for 30 min with a new solution only containing DMSO, S1P or FTY720P. Fluorescent transferrin-Alexa Fluor 647 was used to follow the efficiency of the clathrin-based endocytic pathway. Images obtained using spinning disc confocal microscopy correspond to a center plane from which quantification data was obtained. An acid wash step at the end of the experiment was used to remove the surface bound transferrin and anti-HA antibodies. Scale bar, 10 μm.

Article Snippet: Reagents Sphingosine-1-phosphate (S1P) and FTY720-phosphate (FTY720P) were from Echelon Bioscience (Salt Lake City, UT), poly-d-lysine (Sigma) and collagen (BD Biosciences).

Techniques: Expressing, Incubation, Modification, Confocal Microscopy

Journal: Experimental and Therapeutic Medicine

Article Title: Combination of C-X-C motif chemokine 9 and C-X-C motif chemokine 10 antibodies with FTY720 prolongs the survival of cardiac retransplantation allografts in a mouse model

doi: 10.3892/etm.2015.2204

Figure Lengend Snippet: Post-transplantation treatment regimens.

Article Snippet: Similarly, FTY720 (R&D Systems, Inc.) was administered (0.2 mg; ip) to the transplant recipients on days −1, +1, +3 and every two days until rejection.

Techniques: Control

Journal: Experimental and Therapeutic Medicine

Article Title: Combination of C-X-C motif chemokine 9 and C-X-C motif chemokine 10 antibodies with FTY720 prolongs the survival of cardiac retransplantation allografts in a mouse model

doi: 10.3892/etm.2015.2204

Figure Lengend Snippet: Cardiac allograft survival time in transplant recipient mice. C57BL/6 mice received BALB/c cardiac allografts and were divided into four groups: Control, CXCL9 Ab and CXCL10 Ab, FTY720, and combined (n=6 in each group). The graft median survival time was 3.5, 4.7, 4.7 and 9.3 days in each group, respectively. Combined group, CXCL9 Ab + CXCL10 Ab + FTY720; CXCL, C-X-C motif chemokine; Ab, antibody.

Article Snippet: Similarly, FTY720 (R&D Systems, Inc.) was administered (0.2 mg; ip) to the transplant recipients on days −1, +1, +3 and every two days until rejection.

Techniques: Control

Journal: Experimental and Therapeutic Medicine

Article Title: Combination of C-X-C motif chemokine 9 and C-X-C motif chemokine 10 antibodies with FTY720 prolongs the survival of cardiac retransplantation allografts in a mouse model

doi: 10.3892/etm.2015.2204

Figure Lengend Snippet: Graft pathological sections of each experimental group showing the rejection of cardiac allografts. On day 4 after heart transplantation, grafts were harvested and prepared for histological evaluation. (A and B) Control group at (A) ×100 and (B) ×400 magnification (ISHLT grade 4); (C and D) CXCL9 Ab and CXCL10 Ab group at (C) ×100 and (D) ×400 magnification (ISHLT grade 3A); (E and F) FTY720 group at (E) ×100 and (F) ×400 magnification (ISHLT grade 3B); and (G and H) combined group at (G) ×100 and (H) ×400 magnification (ISHLT grade 1A). Combined group, CXCL9 Ab + CXCL10 Ab + FTY720; ISHLT, International Society for Heart and Lung Transplantation; CXCL, C-X-C motif chemokine; Ab, antibody.

Article Snippet: Similarly, FTY720 (R&D Systems, Inc.) was administered (0.2 mg; ip) to the transplant recipients on days −1, +1, +3 and every two days until rejection.

Techniques: Transplantation Assay, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Combination of C-X-C motif chemokine 9 and C-X-C motif chemokine 10 antibodies with FTY720 prolongs the survival of cardiac retransplantation allografts in a mouse model

doi: 10.3892/etm.2015.2204

Figure Lengend Snippet: Relative gene expression levels of IL-2, IL-10, IFN-γ and TGF-β in the cardiac allografts of each group. Allograft mRNA was analyzed using qPCR. The gene expression level of IL-2 was significantly downregulated, while the gene expression level of TGF-β was significantly upregulated in the experimental groups compared with the control group. * P<0.05, ** P<0.01 and *** P<0.001 vs. the control group. Combined group, CXCL9 Ab + CXCL10 Ab + FTY720; CXCL, C-X-C motif chemokine; Ab, antibody; IL, interleukin; IFN, interferon; TGF, transforming growth factor; qPCR, quantitative polymerase chain reaction.

Article Snippet: Similarly, FTY720 (R&D Systems, Inc.) was administered (0.2 mg; ip) to the transplant recipients on days −1, +1, +3 and every two days until rejection.

Techniques: Gene Expression, Control, Real-time Polymerase Chain Reaction

Journal: Experimental and Therapeutic Medicine

Article Title: Combination of C-X-C motif chemokine 9 and C-X-C motif chemokine 10 antibodies with FTY720 prolongs the survival of cardiac retransplantation allografts in a mouse model

doi: 10.3892/etm.2015.2204

Figure Lengend Snippet: Serum concentration levels of (A) IL-2, (B) IL-10, (C) IFN-γ and (D) TGF-β, in the peripheral blood of recipient mice. The concentrations of IL-2 and IFN-γ were lower in the experimental groups than those in the control group. The concentration of IL-10 was higher in the serum of the combined experimental group than that in the control. ** P<0.01 and *** P<0.001 vs. the control group. Combined group, CXCL9 Ab + CXCL10 Ab + FTY720; CXCL, C-X-C motif chemokine; Ab, antibody; IL, interleukin; IFN, interferon; TGF, transforming growth factor.

Article Snippet: Similarly, FTY720 (R&D Systems, Inc.) was administered (0.2 mg; ip) to the transplant recipients on days −1, +1, +3 and every two days until rejection.

Techniques: Concentration Assay, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Combination of C-X-C motif chemokine 9 and C-X-C motif chemokine 10 antibodies with FTY720 prolongs the survival of cardiac retransplantation allografts in a mouse model

doi: 10.3892/etm.2015.2204

Figure Lengend Snippet: Effect of drug treatment on T-cell proliferation. The proliferative response in each group was assessed with the mixed lymphocyte reaction. ** P<0.01 and *** P<0.001 vs. the control group. Combined group, CXCL9 Ab + CXCL10 Ab + FTY720; CXCL, C-X-C motif chemokine; Ab, antibody; CD, cluster of differentiation.

Article Snippet: Similarly, FTY720 (R&D Systems, Inc.) was administered (0.2 mg; ip) to the transplant recipients on days −1, +1, +3 and every two days until rejection.

Techniques: Control

Journal: Journal of Neuroinflammation

Article Title: Increased levels of sphingosine-1-phosphate in cerebrospinal fluid of patients diagnosed with tick-borne encephalitis

doi: 10.1186/s12974-014-0193-4

Figure Lengend Snippet: F-actin structure in rat astrocytes (confluent culture) treated with S1P, recombinant human plasma gelsolin and FTY720P, or their combination. At 8 hours incubation the cells were fixed with 4% paraformaldehyde, permeabilized with Triton X-100 and F-actin was stained with Phalloidin-FITC. Arrows indicate gaps in astrocyte monolayers. Data from one representative experiment are shown (A) . B shows quantitative analysis of F-actin fluorescence in the astrocyte monolayer under treatment, indicated by numbers 1 to 10 in the left lower corner of each picture. *Significantly different. Error bars represent standard error of the mean. S1P – sphingosine – 1– phosphate, GSN – recombinant human plasma gelsolin.

Article Snippet: S1P (S9666) was purchased from Sigma Aldrich (St Louis, Missouri, United States). (S)- fingolimod phosphate (FTY720P; B-0721) was purchased from Echelon Biosciences Inc. (Salt Lake City, Utah, United States).

Techniques: Recombinant, Incubation, Staining, Fluorescence

Journal: Journal of Neuroinflammation

Article Title: Increased levels of sphingosine-1-phosphate in cerebrospinal fluid of patients diagnosed with tick-borne encephalitis

doi: 10.1186/s12974-014-0193-4

Figure Lengend Snippet: S1P, recombinant human plasma gelsolin (rhGSN) and FTY720P prevent release of IL-6 from rat astrocytes. IL-6 release from rat astrocytes 8 hours after addition of S1P, (rhGSN and FTY720P (A) . Decrease of IL-6 release from rat astrocytes activated with S1P (5 μM) in the presence of rhGSN (5 μM) and FTY720P (5 μM) (B) . *Significantly different. Error bars represent standard deviation of the mean. IL–6 – interleukin 6, S1P – shingosine-1-phosphate

Article Snippet: S1P (S9666) was purchased from Sigma Aldrich (St Louis, Missouri, United States). (S)- fingolimod phosphate (FTY720P; B-0721) was purchased from Echelon Biosciences Inc. (Salt Lake City, Utah, United States).

Techniques: Recombinant, Standard Deviation

Journal: ACS Nano

Article Title: Enhancing Lung Recovery: Inhaled Poly(lactic- co -glycolic) Acid Encapsulating FTY720 and Nobiletin for Lipopolysaccharide-Induced Lung Injury, with Advanced Inhalation Tower Technology

doi: 10.1021/acsnano.3c12532

Figure Lengend Snippet: (A) The number distribution of coencapsulated FTY720 and NOB NPs was prepared by the emulsification method using PLGA. (B) The morphology of the FTY720-NOB-PLGA NPs with a field of view of 30K.

Article Snippet: FTY720 was purchased from TargetMol Chemicals Inc. (USA).

Techniques: Emulsification

Journal: ACS Nano

Article Title: Enhancing Lung Recovery: Inhaled Poly(lactic- co -glycolic) Acid Encapsulating FTY720 and Nobiletin for Lipopolysaccharide-Induced Lung Injury, with Advanced Inhalation Tower Technology

doi: 10.1021/acsnano.3c12532

Figure Lengend Snippet: (A) FTIR spectra of the (a) FTY720, (b) NOB, (c) PLGA, and (d) FTY720-NOB-PLGA NPs with wave numbers from 500 to 4000 cm –1 . The FTY720-NOB-PLGA NPs underwent drug release at 1× PBST and 37 °C. Concentrations of FTY720 and NOB were determined at wavelengths of 200 and 330 nm and expressed as (B) grams of cumulative drug and (C) percent of total cumulative drug.

Article Snippet: FTY720 was purchased from TargetMol Chemicals Inc. (USA).

Techniques:

Journal: ACS Nano

Article Title: Enhancing Lung Recovery: Inhaled Poly(lactic- co -glycolic) Acid Encapsulating FTY720 and Nobiletin for Lipopolysaccharide-Induced Lung Injury, with Advanced Inhalation Tower Technology

doi: 10.1021/acsnano.3c12532

Figure Lengend Snippet: (A) Schematic diagram illustrating the design of the animal model. (B) Evaluation of IL-6 and TNF-α secretion in bronchoalveolar lavage fluid (BALF) from LPS-induced lung injury mice ( n = 3 per group) after 24 h of treatment with PBS, NOB-PLGA NPs, and FTY720-NOB-PLGA NPs using an ELISA kit. (C) Histopathological examination of lung tissues in mice ( n = 3 per group) with LPS-induced lung injury treated with PBS, NOB-PLGA NPs, and FTY720-NOB-PLGA NPs at 24 h, displayed at an original magnification of 100×. (D) Panoramic images of NF-κB obtained through the TissueFAXs platform and quantification of NF-κB expression percentage in the entire lung section (*** p < 0.001, ** p < 0.01, and * p < 0.05).

Article Snippet: FTY720 was purchased from TargetMol Chemicals Inc. (USA).

Techniques: Animal Model, Enzyme-linked Immunosorbent Assay, Expressing

Journal: ACS Nano

Article Title: Enhancing Lung Recovery: Inhaled Poly(lactic- co -glycolic) Acid Encapsulating FTY720 and Nobiletin for Lipopolysaccharide-Induced Lung Injury, with Advanced Inhalation Tower Technology

doi: 10.1021/acsnano.3c12532

Figure Lengend Snippet: (A) Schematic diagram of the animal model examining the endurance of FTY720-NOB-PLGA NPs’ efficacy over a 48 h period. (B) Assessment of IL-6 and TNF-α secretion in bronchoalveolar lavage fluid (BALF) from LPS-induced lung injury mice ( n = 3 per group) 48 h post-treatment with PBS, NOB-PLGA NPs, and FTY720-NOB-PLGA NPs using an ELISA kit. (C) Lung pathology evaluation in mice ( n = 3 per group) with LPS-induced lung injury treated with different drugs at 48 h, displayed at an original magnification of 100×. (D) Panoramic images of NF-κB obtained through the TissueFAXs platform, along with quantification of NF-κB expression percentage in the entire lung section (*** p < 0.001, ** p < 0.01, and * p < 0.05).

Article Snippet: FTY720 was purchased from TargetMol Chemicals Inc. (USA).

Techniques: Animal Model, Enzyme-linked Immunosorbent Assay, Expressing

Journal: ACS Nano

Article Title: Enhancing Lung Recovery: Inhaled Poly(lactic- co -glycolic) Acid Encapsulating FTY720 and Nobiletin for Lipopolysaccharide-Induced Lung Injury, with Advanced Inhalation Tower Technology

doi: 10.1021/acsnano.3c12532

Figure Lengend Snippet: (A) Schematic illustration of FTY720-NOB-PLGA NPs inhalation for 30 min daily over three consecutive days, followed by sacrifice at 96 h. (B) Comparison of the impact of different treatments (PBS and FTY720-NOB-PLGA NPs; 30 min/day for 3 days) on IL-6 and TNF-α secretion in BALF using an ELISA kit. (C) Lung pathology assessment in mice ( n = 3 per group) with LPS-induced lung injury treated with different drugs for three consecutive days. (D) Panoramic NF-κB images obtained through the TissueFAXs platform, and quantification of NF-κB expression percentage in the entire lung section. Mean ± SD values are presented, and p -values were determined using the Student’s t test (*** p < 0.001, ** p < 0.01, and * p < 0.05).

Article Snippet: FTY720 was purchased from TargetMol Chemicals Inc. (USA).

Techniques: Comparison, Enzyme-linked Immunosorbent Assay, Expressing

Journal: ACS Nano

Article Title: Enhancing Lung Recovery: Inhaled Poly(lactic- co -glycolic) Acid Encapsulating FTY720 and Nobiletin for Lipopolysaccharide-Induced Lung Injury, with Advanced Inhalation Tower Technology

doi: 10.1021/acsnano.3c12532

Figure Lengend Snippet: Histological analysis of lung tissue sections from control, LPS-induced acute lung injury, and post-treatment with FTY720-NOB-PLGA NPs. (A) Tissue sections were stained using H&E and immunohistochemical staining for MPO, CD68, and CD3. Positive cells are brown (magnification, 400×). Scale bar, 10 μm. (B) Quantification of neutrophils (MPO), macrophages (CD68), and T cells (CD3) per high-power field (HPF) derived from IHC data ( n = 3 per group). Data are presented as mean ± SD. Statistical significance was assessed using the Student’s t test (*** p < 0.001, ** p < 0.01).

Article Snippet: FTY720 was purchased from TargetMol Chemicals Inc. (USA).

Techniques: Control, Staining, Immunohistochemical staining, Derivative Assay

Journal: ACS Nano

Article Title: Enhancing Lung Recovery: Inhaled Poly(lactic- co -glycolic) Acid Encapsulating FTY720 and Nobiletin for Lipopolysaccharide-Induced Lung Injury, with Advanced Inhalation Tower Technology

doi: 10.1021/acsnano.3c12532

Figure Lengend Snippet: Multiplexed mIF assay of immune cell infiltration. (A) Representative images from the mIF assay on lung tissue from mice post-LPS stimulation with or without FTY720-NOB-PLGA nanoparticle inhalation treatment, showing staining for MPO (neutrophils), CD68 (macrophages), CD3 (T cells), and DAPI (nuclei). Scale bar, 100 μm. (B) The bar graph illustrates the proportion of neutrophils, macrophages, and T cells infiltrating the lungs of mice following LPS stimulation with or without FTY720-NOB-PLGA nanoparticle inhalation treatment. Data are presented as mean ± SD, calculated from four fields per mouse lung. Statistical significance was determined using the Student’s t test (*** p < 0.001).

Article Snippet: FTY720 was purchased from TargetMol Chemicals Inc. (USA).

Techniques: Staining

Journal: ACS Nano

Article Title: Enhancing Lung Recovery: Inhaled Poly(lactic- co -glycolic) Acid Encapsulating FTY720 and Nobiletin for Lipopolysaccharide-Induced Lung Injury, with Advanced Inhalation Tower Technology

doi: 10.1021/acsnano.3c12532

Figure Lengend Snippet: Body weight, organ function, and respiratory rate in control group and treatment group. (A) Body weight percentage of mice in response to LPS and FTY720-NOB-PLGA NPs. The values presented are the mean ± SD. p values were obtained using the Student’s t test (*** p < 0.001). (B) Blood biochemical results of the FTY720-NOB-PLGA NPs following inhalation into mice ( n = 5). The results show mean and standard deviation of alanine aminotransferase (ALT), creatinine (CRE), total bilirubin (TBIL), and blood urea nitrogen (BUN). (C) Respiratory parameters: Mean of breathing frequency was analyzed during day 1 to day 3 through plethysmography associated with the Allay restrainer.

Article Snippet: FTY720 was purchased from TargetMol Chemicals Inc. (USA).

Techniques: Control, Standard Deviation

Journal: Scientific reports

Article Title: Effect of FTY720P on lipid accumulation in HEPG2 cells.

doi: 10.1038/s41598-023-46011-4

Figure Lengend Snippet: Figure 2. S1P and FTY720P induce lipid accumulation in HepG2 cells. Lipid accumulation in the presence of (a) Sphk1 inhibitor PF543; (b) blockers of all S1PRs. Values are means ± SEM. N = 4. Bars not sharing a common letter are considered significantly different from each other at P < 0.05; c) various concentrations of the S1P analog FTY720P. Values are means ± SEM; N = 3. *Significantly different from the control at P < 0.05.

Article Snippet: FTY720P (Cat # sc-205332A), rosiglitazone (Cat # sc-202795), anti-SREBP1 (Cat# sc-365513, Lot#B1821), anti-p-mTOR (Cat# sc-293132, Lot#K2717), anti-mTOR (Cat# sc-8319, Lot#K2415), anti-GAPDH (Cat# sc-47724, Lot# H0917) mouse primary antibodies as well as anti-mouse horse radish peroxidase (HRP) conjugated secondary antibodies (Cat# sc-2005, Lot# C2011) were purchased all from Santa Cruz Biotechnology, CA, USA.

Techniques: Control

Journal: Scientific reports

Article Title: Effect of FTY720P on lipid accumulation in HEPG2 cells.

doi: 10.1038/s41598-023-46011-4

Figure Lengend Snippet: Figure 3. Effect of FTY720P on lipid accumulation in the presence of (a) S1PR1 blocker W146. (b) S1PR2 blocker JTE 013. (c) S1PR4 blocker CYM50358. Values are means ± SEM; N = 5. Bars not sharing a common letter are considered significantly different from each other at P < 0.05.

Article Snippet: FTY720P (Cat # sc-205332A), rosiglitazone (Cat # sc-202795), anti-SREBP1 (Cat# sc-365513, Lot#B1821), anti-p-mTOR (Cat# sc-293132, Lot#K2717), anti-mTOR (Cat# sc-8319, Lot#K2415), anti-GAPDH (Cat# sc-47724, Lot# H0917) mouse primary antibodies as well as anti-mouse horse radish peroxidase (HRP) conjugated secondary antibodies (Cat# sc-2005, Lot# C2011) were purchased all from Santa Cruz Biotechnology, CA, USA.

Techniques:

Journal: Scientific reports

Article Title: Effect of FTY720P on lipid accumulation in HEPG2 cells.

doi: 10.1038/s41598-023-46011-4

Figure Lengend Snippet: Figure 4. S1PR3 and Gq mediate the FTY720P effect on lipid accumulation. (a) Effect of FTY720P on lipid accumulation in the presence of S1PR3 blocker CAY10444. (b) Effect of FTY720P on lipid accumulation in the presence of the Gq inhibitor YM254890. (c) Protein expression of S1PR3. The blot is representative of an experiment repeated 3 times. Values were normalized to GAPDH and reported as arbitrary densitometry units. All values are means ± SEM; N = 3. Bars not sharing a common letter are considered significantly different from each other at P < 0.05.

Article Snippet: FTY720P (Cat # sc-205332A), rosiglitazone (Cat # sc-202795), anti-SREBP1 (Cat# sc-365513, Lot#B1821), anti-p-mTOR (Cat# sc-293132, Lot#K2717), anti-mTOR (Cat# sc-8319, Lot#K2415), anti-GAPDH (Cat# sc-47724, Lot# H0917) mouse primary antibodies as well as anti-mouse horse radish peroxidase (HRP) conjugated secondary antibodies (Cat# sc-2005, Lot# C2011) were purchased all from Santa Cruz Biotechnology, CA, USA.

Techniques: Expressing

Journal: Scientific reports

Article Title: Effect of FTY720P on lipid accumulation in HEPG2 cells.

doi: 10.1038/s41598-023-46011-4

Figure Lengend Snippet: Figure 5. Effect of FTY720P on lipid accumulation in the presence of (a) the PI3K inhibitor wortmannin. (b) the mTOR inhibitor rapamycin. All values are means ± SEM; N = 4. Bars not sharing a common letter are considered significantly different from each other at P < 0.05.

Article Snippet: FTY720P (Cat # sc-205332A), rosiglitazone (Cat # sc-202795), anti-SREBP1 (Cat# sc-365513, Lot#B1821), anti-p-mTOR (Cat# sc-293132, Lot#K2717), anti-mTOR (Cat# sc-8319, Lot#K2415), anti-GAPDH (Cat# sc-47724, Lot# H0917) mouse primary antibodies as well as anti-mouse horse radish peroxidase (HRP) conjugated secondary antibodies (Cat# sc-2005, Lot# C2011) were purchased all from Santa Cruz Biotechnology, CA, USA.

Techniques:

Journal: Scientific reports

Article Title: Effect of FTY720P on lipid accumulation in HEPG2 cells.

doi: 10.1038/s41598-023-46011-4

Figure Lengend Snippet: Figure 6. Effect of FTY720P on lipid accumulation in the presence of (a) the SREBP inhibitor fatostatin. (b) The PPARγ inhibitor GW9662. Values are means ± SEM; N = 5. Bars not sharing a common letter are considered significantly different from each other at P < 0.05.

Article Snippet: FTY720P (Cat # sc-205332A), rosiglitazone (Cat # sc-202795), anti-SREBP1 (Cat# sc-365513, Lot#B1821), anti-p-mTOR (Cat# sc-293132, Lot#K2717), anti-mTOR (Cat# sc-8319, Lot#K2415), anti-GAPDH (Cat# sc-47724, Lot# H0917) mouse primary antibodies as well as anti-mouse horse radish peroxidase (HRP) conjugated secondary antibodies (Cat# sc-2005, Lot# C2011) were purchased all from Santa Cruz Biotechnology, CA, USA.

Techniques:

Journal: Scientific reports

Article Title: Effect of FTY720P on lipid accumulation in HEPG2 cells.

doi: 10.1038/s41598-023-46011-4

Figure Lengend Snippet: Figure 13. The signaling pathway activated by FTY720P.

Article Snippet: FTY720P (Cat # sc-205332A), rosiglitazone (Cat # sc-202795), anti-SREBP1 (Cat# sc-365513, Lot#B1821), anti-p-mTOR (Cat# sc-293132, Lot#K2717), anti-mTOR (Cat# sc-8319, Lot#K2415), anti-GAPDH (Cat# sc-47724, Lot# H0917) mouse primary antibodies as well as anti-mouse horse radish peroxidase (HRP) conjugated secondary antibodies (Cat# sc-2005, Lot# C2011) were purchased all from Santa Cruz Biotechnology, CA, USA.

Techniques:

Journal: Journal of Leukocyte Biology

Article Title: Rapid externalization of 27-kDa heat shock protein (HSP27) and atypical cell death in neutrophils treated with the sphingolipid analog drug FTY720

doi: 10.1189/jlb.3vma1114-522rr

Figure Lengend Snippet: Figure 5. RIP1K and HSP90 are involved in neutrophil death induced by FTY720. (A and B) Flow cytometric quantification of Annexin V+PI+, Annexin V+PI2, and HSP27+ neutrophils after preincubation or not with necrostatin-1 (20 mM) or Z-IETD-FMK (20 mM) for 30 min and treatment or not with FTY720 (10 mM) for 3 h. Data are from 7 independent experiments. *P , 0.05; **P , 0.01; ***P , 0.001. (C and D) Neutrophil lysates analyzed by Western blotting for caspase-8 cleavage or MLKL phosphorylation after cell death induc- tion by FTY720. Where indicated, Q-VD-OPh (20 mM), necrostatin-1 (20 mM), DPI (20 mM), or NSA (5 mM) was added for 30 min before stimulation. GAPDH analysis was performed as loading control. The results of 1 representative experiment out of 4 are presented. (E) Flow cytometric quantification of HSP27+ neutrophils incubated for 30 min in the presence of geldanamycin (10 mM), DMAG (20 mM), or radicicol (10 mM) before treatment with FTY720 (10 mM) for 3 h. Data are from 7 independent experiments. *P , 0.05; **P , 0.01.

Article Snippet: Cells were incubated with FTY720 and FTY720 (S)-phosphate (both from Santa Cruz Biotechnology, Santa Cruz, CA, USA) or cycloheximide (Sigma, St. Louis, MO, USA) at 3 3 10/ml for 3 h. In some cases, neutrophils were preincubated for 30 min with necrostatin-1 or Z-IETD-FMK (both from Sigma); Q-VD-OPh (R&D Systems, Minneapolis, MN, USA); NSA (Tocris, Bristol, United Kingdom); geldanamycin, 17-DMAG, and radicicol (all from InvivoGen, San Diego, CA, USA); OA (Santa Cruz Biotechnology); or DPI (Sigma).

Techniques: Western Blot, Phospho-proteomics, Control, Incubation