Journal: The Journal of allergy and clinical immunology

Article Title: Nasal polyps 15-lipoxygenase 1 promotes CCL26/Eotaxin3 expression through ERK activation

doi: 10.1016/j.jaci.2019.06.037

Figure Lengend Snippet: 15LO1 mRNA and protein are higher in fresh NECs of CRSwNP subjects compared to HCs. Total RNA and protein from freshly brushed NECs were harvested for RT-PCR and Western blot analysis. NP and MT tissues from subjects with CRSwNP and HCs were fixed for Immunohistochemistry studies. (A) 15LO1 mRNA is higher in NECs of NP from CRSwNP subjects compared to HC. (B) 15LO1 protein is higher in NECs of NP from CRSwNP compared to NECs of MT from HCs. (C) Densitometry analysis of 15LO1 protein from CRSwNP subjects and HCs by WB. (D) 15LO1 protein is higher in NECs of NP compared to MT from same CRSwNP subjects. (E) Densitometry analysis of the paired MT and NP 15LO1 protein. (F) Representative immunostaining photomicrograph showed 15LO1 positive staining is upregulated in NECs from CRSwNP subjects than MT of CRSwNP and HCs. Abbreviations: NECs=nasal epithelial cells, HC=healthy controls, NP=nasal polyps, MT=middle turbinate, IT=inferior turbinate, and CRSwNP=chronic rhinosinusitis with nasal polyps.

Article Snippet: Tissue sections were blocked with 5% donkey serum and incubated with primary antibodies against 15LO1 (1: 200 dilution), CCL26 (1:100 dilution), and Cytokeratin-5 (CK5, 1: 200 dilution, Novus, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Immunohistochemistry, Immunostaining, Staining

Journal: The Journal of allergy and clinical immunology

Article Title: Nasal polyps 15-lipoxygenase 1 promotes CCL26/Eotaxin3 expression through ERK activation

doi: 10.1016/j.jaci.2019.06.037

Figure Lengend Snippet: CCL26 mRNA and protein are higher in fresh NECs from CRSwNP compared to HCs and correlated with 15LO1 expression. Total RNA and protein from freshly brushed NECs were harvested for RT-PCR and ELISA analysis. (A) CCL26 mRNA is higher in NECs of NP from CRSwNP subjects compared to HCs. (B) CCL26 mRNA expression correlated with 15LO1 mRNA ex vivo. (C) CCL26 protein (by ELISA) is higher in NECs of MT and NP cell lysates from CRSwNP subjects, especially NP, compared to HCs. (D) CCL26 protein (by ELISA) expression correlates with 15LO1 protein expression (by WB) in NP cell lysates. Abbreviations: NECs=nasal epithelial cells, HC=healthy controls, NP=nasal polyps, MT=middle turbinate, and CRSwNP=chronic rhinosinusitis with nasal polyps.

Article Snippet: Tissue sections were blocked with 5% donkey serum and incubated with primary antibodies against 15LO1 (1: 200 dilution), CCL26 (1:100 dilution), and Cytokeratin-5 (CK5, 1: 200 dilution, Novus, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Ex Vivo

Journal: The Journal of allergy and clinical immunology

Article Title: Nasal polyps 15-lipoxygenase 1 promotes CCL26/Eotaxin3 expression through ERK activation

doi: 10.1016/j.jaci.2019.06.037

Figure Lengend Snippet: Immunostaining results showed that 15LO1/CCL26 and CCL26/cytokeratin-5 positive cell numbers increased in MT and NP tissues from CRSwNP subjects, especially in NP tissues, compared to HCs. NP and MT tissues from subjects with CRSwNP and HCs were fixed for IF analysis. (A) Representative immunostaining photomicrograph showed CCL26 positive cells are expressed in MT and NP epithelial cells of CRSwNP subjects. CCL26 colocalized with 15LO1 in MT (n=6) and NP (n=6) epithelial cells of CRSwNP. (B) Representative immunostaining photomicrograph showed no positive 15LO1/CCL26 double staining in MT (n=5) tissues of HCs. (C) Quantitation of 15LO1/CCL26 positive signals in NP and MT epithelial cells from CRSwNP and HCs. (D) Representative immunostaining photomicrograph showed CCL26 colocalized with cytokeratin-5 in MT (n=3) and NP (n=3) epithelial cells from CRSwNP subjects. (E) Quantitation of CCL26/CK5 positive signals in NP and MT tissues from CRSwNP and HCs (n=3). Abbreviations: HC=healthy controls, NP=nasal polyps, MT=middle turbinate, and CRSwNP=chronic rhinosinusitis with nasal polyps.

Article Snippet: Tissue sections were blocked with 5% donkey serum and incubated with primary antibodies against 15LO1 (1: 200 dilution), CCL26 (1:100 dilution), and Cytokeratin-5 (CK5, 1: 200 dilution, Novus, USA).

Techniques: Immunostaining, Double Staining, Quantitation Assay

Journal: The Journal of allergy and clinical immunology

Article Title: Nasal polyps 15-lipoxygenase 1 promotes CCL26/Eotaxin3 expression through ERK activation

doi: 10.1016/j.jaci.2019.06.037

Figure Lengend Snippet: IL-13 induces 15LO1 and CCL26 expression in ALI cultured NECs and 15LO1 positively correlates with CCL26 expression in vitro. ALI cultured primary NECs were stimulated with IL-13 for 5 days. Cells were harvested for RT-PCR and Western blot analysis. Lower supernatants were harvested for CCL26 levels by ELISA. IL-13 increased (A) 15LO1 mRNA expression, (B) CCL26 mRNA expression and (C) 15LO1 mRNA correlates with CCL26 mRNA expression in vitro. (D) IL-13 induces intracellular 15LO1 and CCL26 protein in cell lysates. (E) IL-13 induced 15LO1 protein correlates with CCL26 protein in vitro. Abbreviations: ALI=air-liquid interface, NECs=nasal epithelial cells.

Article Snippet: Tissue sections were blocked with 5% donkey serum and incubated with primary antibodies against 15LO1 (1: 200 dilution), CCL26 (1:100 dilution), and Cytokeratin-5 (CK5, 1: 200 dilution, Novus, USA).

Techniques: Expressing, Cell Culture, In Vitro, Reverse Transcription Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: The Journal of allergy and clinical immunology

Article Title: Nasal polyps 15-lipoxygenase 1 promotes CCL26/Eotaxin3 expression through ERK activation

doi: 10.1016/j.jaci.2019.06.037

Figure Lengend Snippet: ALOX15 DsiRNA and 15LO1 specific inhibitor BLX2477 decreased IL-13 induced CCL26 expression. ALI cultured NECs were treated with ALOX15 DsiRNA for 5 days in the presence or absence of IL-13. ALI cultured NECs were treated with BLX2477 for 3 days in the presence of IL-13. Cells were harvested for RT-PCR and Western blot. Lower supernatants were harvested for CCL26 protein by ELISA. 15LO1 knockdown decreases (A) intracellular CCL26 mRNA and (B) intracellular CCL26 protein and (C) secreted CCL26 protein in lower supernatants. (D) Densitometry analysis of intracellular CCL26 protein in cell lysates. (E) The 15LO1 inhibitor decreases IL-13 induced CCL26 mRNA, (F) and CCL26 protein in cell lysates and (G) secreted CCL26 protein in lower supernatants. (H) Densitometry analysis of CCL26 protein in cell lysates. Abbreviations: ALI=air-liquid interface, NECs=nasal epithelial cells.

Article Snippet: Tissue sections were blocked with 5% donkey serum and incubated with primary antibodies against 15LO1 (1: 200 dilution), CCL26 (1:100 dilution), and Cytokeratin-5 (CK5, 1: 200 dilution, Novus, USA).

Techniques: Expressing, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay, Knockdown

Journal: The Journal of allergy and clinical immunology

Article Title: Nasal polyps 15-lipoxygenase 1 promotes CCL26/Eotaxin3 expression through ERK activation

doi: 10.1016/j.jaci.2019.06.037

Figure Lengend Snippet: ALOX15 DsiRNA and 15LO1 inhibitor BLX2477 decreased pERK expression. Blocking ERK pathway inhibits CCL26 production in ALI cultured NECs. (A) ALOX15 DsiRNA decreased pERK but not tERK by WB. (B) The 15LO1 specific inhibitor decreased pERK but not tERK by WB. (C) ALOX15 DsiRNA and 15LO1 specific inhibitor BLX2477 decreased pERK/tERK ratio. (D) ERK inhibitor PD98059 decreased pERK and CCL26 protein by WB. (E) ERK inhibitor U0126 decreased pERK and CCL26 protein by WB. (F) ERK pathway inhibitors PD98059 and U0126 decreased CCL26 mRNA in ALI cultured NECs. (G) The ERK pathway inhibitor PD98059 and U0126 decreased secreted CCL26 in supernatants. Abbreviations: ALI=air-liquid interface, NECs=nasal epithelial cells, WB=western blot.

Article Snippet: Tissue sections were blocked with 5% donkey serum and incubated with primary antibodies against 15LO1 (1: 200 dilution), CCL26 (1:100 dilution), and Cytokeratin-5 (CK5, 1: 200 dilution, Novus, USA).

Techniques: Expressing, Blocking Assay, Cell Culture, Western Blot

Journal: The Journal of allergy and clinical immunology

Article Title: Nasal polyps 15-lipoxygenase 1 promotes CCL26/Eotaxin3 expression through ERK activation

doi: 10.1016/j.jaci.2019.06.037

Figure Lengend Snippet: Increased pERK correlates with 15LO1 and CCL26 in fresh NECs. (A) Phosphorylated ERK is increased in fresh NECs of NP compared to HCs. (B) Densitometry analysis of pERK/tERK in fresh NECs of CRSwNP and HCs. (C) pERK/tERK ratio correlates with 15LO1 protein in fresh NECs of CRSwNP and HCs. (D) pERK/tERK ratio correlates with CCL26 protein (by ELISA) in fresh NECs of CRSwNP. Abbreviations: NECs=nasal epithelial cells, HC=healthy controls, NP=nasal polyps, MT=middle turbinate, and CRSwNP=chronic rhinosinusitis with nasal polyps.

Article Snippet: Tissue sections were blocked with 5% donkey serum and incubated with primary antibodies against 15LO1 (1: 200 dilution), CCL26 (1:100 dilution), and Cytokeratin-5 (CK5, 1: 200 dilution, Novus, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Cancer Science

Article Title: 15‐Lipoxygenase‐1 expression suppresses the invasive properties of colorectal carcinoma cell lines HCT‐116 and HT‐29

doi: 10.1111/j.1349-7006.2009.01313.x

Figure Lengend Snippet: 15‐Lipoxygenase‐1 (15‐LO‐1) overexpression in HCT‐116 and HT‐29 colorectal carcinoma cell lines. (A) RT‐PCR analysis of the 15‐LO‐1 transcript. For HCT‐116 cells: lane 1, 15‐LO‐1 vector (positive control); lane 2, 15‐LO‐1 expressing clone 1E7; lane 3, 15‐LO‐1 expressing clone 1F8; lane 4, empty vector (pcDNA3.1) transfected cells; lane 5, parental HCT‐116 cells; lane 6, negative control. For HT‐29 cells transiently transfected with a 15‐LO‐1 vector: lane 1, positive control; lane 2, 15‐LO‐1 expressing cells; lane 3, empty vector transfected cells; lane 4, mock transfected cells; lane 5, untransfected parental HT‐29 cells; lane 6, negative control. Band intensities were normalized using GAPDH. (B) Western blot analysis of 15‐LO‐1 protein expression. For HCT‐116: lane 1, 15‐LO‐1 expressing clone 1E7; lane 2, 15‐LO‐1 expressing clone 1F8; lane 3, empty vector transfected cells; lane 4, untransfected parental cells. For HT‐29 cells: lane 1, 15‐LO‐1 transfected cells; lane 2, empty vector transfected cells; lane 3, parental untransfected cells. Equal protein loading was confirmed by probing with a GAPDH antibody. (C) The enzymatic activity of 15‐LO‐1 transfected HCT‐116 (HCT116_15LO1, upper panel) and HT‐29 (HT29_15LO1, lower panel) cells was determined from their content of 13(S)‐HODE by an ELISA assay. A decrease in the enzymatic formation of 13(S)‐HODE was observed when HCT‐116 cells expressing 15‐LO‐1 were incubated with PD146176 (1 μm) for 24 h (15LO1 + PD, upper panel). *P < 0.05 (HCT‐116) and ***P < 0.001 (HT‐29) reflect significantly more 13(S)‐HODE compared to the empty vector (EV) expressing cells (HCT116_EV and HT29_EV). Error bars represent the SD of five independent experiments. ns, not significant.

Article Snippet: The membranes were blocked in 5–10% skim milk and were incubated overnight with the appropriate primary antibodies: 15‐LO‐1 (1:1000 dilution; Cayman Chemical, Ann Arbor, MI, USA); XIAP (1:500 dilution); or MTA‐1 (1:150 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA); followed by incubation for 1 h with a HRP‐conjugated goat anti‐rabbit (1:3300), rabbit anti‐sheep (1:2000) or goat anti‐mouse (1:2000) secondary antibody.

Techniques: Over Expression, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation, Positive Control, Expressing, Transfection, Negative Control, Western Blot, Activity Assay, Enzyme-linked Immunosorbent Assay, Incubation

Journal: Cancer Science

Article Title: 15‐Lipoxygenase‐1 expression suppresses the invasive properties of colorectal carcinoma cell lines HCT‐116 and HT‐29

doi: 10.1111/j.1349-7006.2009.01313.x

Figure Lengend Snippet: 15‐Lipoxygenase‐1 (15‐LO‐1) reduces the proliferation of colon cancer cells. (A) Two clones of HCT‐116 cells stably expressing 15‐LO‐1 (1E7 and 1F8) were plated in a 96‐well plate and allow to grow for 96 h. The cellular proliferation was determined every 24 h by an MTT assay. The 15‐LO‐1 expressing cells proliferated significantly more slowly (**P < 0.01; ***P < 0.001) compared to empty vector (EV) transfected cells. (B) HT‐29 cells were seeded in 96‐well plates and transiently transfected with the 15‐LO‐1 vector. The proliferation was determined 48 h later by an MTT assay. Significantly slower growth (***P < 0.001) was observed when these cells were transfected with 15‐LO‐1 compared to empty pcDNA3.1 transfected, mock transfected, and parental untransfected HT‐29 cells (HT29_UT). Error bars represent the SD of three independent experiments. ns, not significant.

Article Snippet: The membranes were blocked in 5–10% skim milk and were incubated overnight with the appropriate primary antibodies: 15‐LO‐1 (1:1000 dilution; Cayman Chemical, Ann Arbor, MI, USA); XIAP (1:500 dilution); or MTA‐1 (1:150 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA); followed by incubation for 1 h with a HRP‐conjugated goat anti‐rabbit (1:3300), rabbit anti‐sheep (1:2000) or goat anti‐mouse (1:2000) secondary antibody.

Techniques: Clone Assay, Stable Transfection, Expressing, MTT Assay, Plasmid Preparation, Transfection

Journal: Cancer Science

Article Title: 15‐Lipoxygenase‐1 expression suppresses the invasive properties of colorectal carcinoma cell lines HCT‐116 and HT‐29

doi: 10.1111/j.1349-7006.2009.01313.x

Figure Lengend Snippet: 15‐Lipoxygenase‐1 (15‐LO‐1) expression induces apoptosis in colon cancer cells. (A) HCT‐116 cells stably expressing 15‐LO‐1 (HCT116_15LO1, upper panel) and HT‐29 cells transiently transfected with 15‐LO‐1 vector (HT29_15LO1, lower panel) as well as control cells were fixed and incubated with acridine orange dye to identify apoptotic cells. Both cell lines had significantly (**P < 0.01 for HCT‐116; *P < 0.05 for HT‐29) greater numbers of apoptotic cells compared to control cells. (B) Western blot analysis of X‐linked inhibitor of apoptosis (XIAP) in 15‐LO‐1 expressing cells (lane 1) showed reduced protein expression compared to EV transfected (lane 2) and parental HCT‐116 and HT‐29 cells (lane 3). Equal protein loading was confirmed by the levels of GAPDH. The numbers in parentheses indicate densitometric values after normalization to GAPDH. (C) Caspase‐3 activity assay carried out with a colorimetric kit indicates significantly higher caspase‐3 activity in HCT‐116 and HT‐29 cells expressing 15‐LO‐1 (*P < 0.05 for all comparisons) compared to control cells. Error bars represent SD of three independent experiments. ns, not significant.

Article Snippet: The membranes were blocked in 5–10% skim milk and were incubated overnight with the appropriate primary antibodies: 15‐LO‐1 (1:1000 dilution; Cayman Chemical, Ann Arbor, MI, USA); XIAP (1:500 dilution); or MTA‐1 (1:150 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA); followed by incubation for 1 h with a HRP‐conjugated goat anti‐rabbit (1:3300), rabbit anti‐sheep (1:2000) or goat anti‐mouse (1:2000) secondary antibody.

Techniques: Expressing, Stable Transfection, Transfection, Plasmid Preparation, Incubation, Western Blot, Caspase-3 Activity Assay, Activity Assay

Journal: Cancer Science

Article Title: 15‐Lipoxygenase‐1 expression suppresses the invasive properties of colorectal carcinoma cell lines HCT‐116 and HT‐29

doi: 10.1111/j.1349-7006.2009.01313.x

Figure Lengend Snippet: 15‐Lipoxygenase‐1 (15‐LO‐1) expression decreases anchorage‐independent growth on soft agar. HCT‐116 colorectal carcinoma cells stably transfected with 15‐LO‐1 vector (HCT116_15LO1) or control cells were grown on 0.6% noble agar for 2 weeks. Colonies were stained with 0.005% crystal violet and counted under a light microscope. The cells expressing 15‐LO‐1 formed significantly fewer colonies (**P < 0.01) compared to empty vector (EV) transfected and parental untransfected (UT) cells (colony formation represented as 100%). Error bars represent three independent experiments carried out in triplicate. ns, not significant. Scale bars, 100 μm.

Article Snippet: The membranes were blocked in 5–10% skim milk and were incubated overnight with the appropriate primary antibodies: 15‐LO‐1 (1:1000 dilution; Cayman Chemical, Ann Arbor, MI, USA); XIAP (1:500 dilution); or MTA‐1 (1:150 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA); followed by incubation for 1 h with a HRP‐conjugated goat anti‐rabbit (1:3300), rabbit anti‐sheep (1:2000) or goat anti‐mouse (1:2000) secondary antibody.

Techniques: Expressing, Stable Transfection, Transfection, Plasmid Preparation, Staining, Light Microscopy

Journal: Cancer Science

Article Title: 15‐Lipoxygenase‐1 expression suppresses the invasive properties of colorectal carcinoma cell lines HCT‐116 and HT‐29

doi: 10.1111/j.1349-7006.2009.01313.x

Figure Lengend Snippet: 15‐Lipoxygenase‐1 (15‐LO‐1) expression causes a reduction in cell motility, represented as wound closing in confluent HCT‐116 colorectal carcinoma cells. Cells expressing 15‐LO‐1 (HCT116_15LO1) were not able to close the wound in the confluent culture, when compared with parental untransfected (HCT116_UT) and empty vector (HCT116_EV) controls, after 72 h. The graphs show 15‐LO‐1 expressing cells have significantly reduced motility (**P < 0.01) with 32% wound closure compared to 56% for EV expressing and 69% for parental HCT‐116 cells. The differences in wound closure between EV transfected and UT cells did not reach statistical significance. Error bars represent SD from three independent experiments each carried out three times. ns, not significant.

Article Snippet: The membranes were blocked in 5–10% skim milk and were incubated overnight with the appropriate primary antibodies: 15‐LO‐1 (1:1000 dilution; Cayman Chemical, Ann Arbor, MI, USA); XIAP (1:500 dilution); or MTA‐1 (1:150 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA); followed by incubation for 1 h with a HRP‐conjugated goat anti‐rabbit (1:3300), rabbit anti‐sheep (1:2000) or goat anti‐mouse (1:2000) secondary antibody.

Techniques: Expressing, Plasmid Preparation, Transfection

Journal: Cancer Science

Article Title: 15‐Lipoxygenase‐1 expression suppresses the invasive properties of colorectal carcinoma cell lines HCT‐116 and HT‐29

doi: 10.1111/j.1349-7006.2009.01313.x

Figure Lengend Snippet: 15‐Lipoxygenase‐1 (15‐LO‐1) expression reduces the ability of HCT‐116 and HT‐29 colorectal carcinoma cells to adhere to the ECM. (A) HCT‐116 cells stably transfected with 15‐LO‐1 or with the empty vector (EV) were seeded in 96‐well plates coated with 50 μg/mL fibronectin and blocked with BSA. The cells were allowed to attach for 2 h then the non‐adherent cells were removed and the MTT assay was carried out to quantify the attached cells at 570 nm. (B) HT‐29 cells transiently transfected for 24 h with the 15‐LO‐1 vector, EV, or mock transfected cells were allowed to attach to the fibronectin coated plates for 2 h. Non‐adherent cell removal and MTT assay were carried out as above. The 15‐LO‐1 expressing cells were found to adhere significantly less (***P < 0.001) to the fibronectin compared to the control cells. Error bars represent SD from two independent experiments carried out in five replicates. ns, not significant.

Article Snippet: The membranes were blocked in 5–10% skim milk and were incubated overnight with the appropriate primary antibodies: 15‐LO‐1 (1:1000 dilution; Cayman Chemical, Ann Arbor, MI, USA); XIAP (1:500 dilution); or MTA‐1 (1:150 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA); followed by incubation for 1 h with a HRP‐conjugated goat anti‐rabbit (1:3300), rabbit anti‐sheep (1:2000) or goat anti‐mouse (1:2000) secondary antibody.

Techniques: Expressing, Stable Transfection, Transfection, Plasmid Preparation, MTT Assay

Journal: Cancer Science

Article Title: 15‐Lipoxygenase‐1 expression suppresses the invasive properties of colorectal carcinoma cell lines HCT‐116 and HT‐29

doi: 10.1111/j.1349-7006.2009.01313.x

Figure Lengend Snippet: 15‐Lipoxygenase‐1 (15‐LO‐1) expression reduces migration and invasion in HCT‐116 colorectal carcinoma cells when compared to empty vector (EV) controls. (A) A Transwell migration assay was done in the presence of serum as a chemoattractant. 15‐LO‐1 transfected HCT‐116 cells migrated through the 8 μm pores of the Transwell in significantly less numbers (**P < 0.01) compared to the control cells. (B) A Transwell invasion assay assaying the ability of the HCT‐116 cells to invade through Matrigel, a reconstituted basement membrane. The photographs were captured in a Leica light microscope under 4× magnification. 15‐LO‐1 expression significantly reduces (**P < 0.01) the ability of HCT‐116 cells to invade through the basement membrane in vitro. The column graph represents the number of cells that could migrate or invade through the Transwell in the experimental and control cell lines. Error bars represent SD from three independent experiments carried out in four replicates. Scale bars, 40 μm.

Article Snippet: The membranes were blocked in 5–10% skim milk and were incubated overnight with the appropriate primary antibodies: 15‐LO‐1 (1:1000 dilution; Cayman Chemical, Ann Arbor, MI, USA); XIAP (1:500 dilution); or MTA‐1 (1:150 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA); followed by incubation for 1 h with a HRP‐conjugated goat anti‐rabbit (1:3300), rabbit anti‐sheep (1:2000) or goat anti‐mouse (1:2000) secondary antibody.

Techniques: Expressing, Migration, Plasmid Preparation, Transwell Migration Assay, Transfection, Transwell Invasion Assay, Membrane, Light Microscopy, In Vitro

Journal: Cancer Science

Article Title: 15‐Lipoxygenase‐1 expression suppresses the invasive properties of colorectal carcinoma cell lines HCT‐116 and HT‐29

doi: 10.1111/j.1349-7006.2009.01313.x

Figure Lengend Snippet: Metastasis associated protein‐1 (MTA‐1) expression is reduced in 15‐lipoxygenase‐1 (15‐LO‐1) expressing colon cancer cells. Total protein lysate isolated from HCT‐116 cells stably expressing 15‐LO‐1 (left) and HT‐29 cells transiently expressing 15‐LO‐1 (right) were probed for MTA‐1 expression using a mouse mAb. A decrease in the protein levels of MTA‐1 is observed in both cell lines in the presence of 15‐LO‐1 expression. For HCT‐116 cells: lane 1, 15‐LO‐1 expressing cells; lane 2, 15‐LO‐1 expressing cells treated with PD146176; lane 3, empty vector expressing cells; lane 4, untransfected parental cells. For HT‐29 cells: lane 1, 15‐LO‐1 expressing cells; lane 2, empty vector expressing cells; lane 3, untransfected parental cells. Equal protein loading was confirmed with the use of GAPDH. The numbers in parentheses for HT‐29 cells indicate densitometric values after normalization to GAPDH.

Article Snippet: The membranes were blocked in 5–10% skim milk and were incubated overnight with the appropriate primary antibodies: 15‐LO‐1 (1:1000 dilution; Cayman Chemical, Ann Arbor, MI, USA); XIAP (1:500 dilution); or MTA‐1 (1:150 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA); followed by incubation for 1 h with a HRP‐conjugated goat anti‐rabbit (1:3300), rabbit anti‐sheep (1:2000) or goat anti‐mouse (1:2000) secondary antibody.

Techniques: Expressing, Isolation, Stable Transfection, Plasmid Preparation

Journal: Cell

Article Title: PEBP1 Wardens Ferroptosis by Enabling Lipoxygenase Generation of Lipid Death Signals

doi: 10.1016/j.cell.2017.09.044

Figure Lengend Snippet: KEY RESOURCE TABLE

Article Snippet: Human 15-LO1 (Human 15-Lipoxygenase) ELISA Kit , MyBioSource , Cat#MBS263359.

Techniques: Produced, Purification, Virus, Recombinant, Saline, Western Blot, Transfection, Protease Inhibitor, Membrane, Enzyme-linked Immunosorbent Assay, Binding Assay, Imaging, Software

Journal:

Article Title: Oxidative Metabolism of Lipoamino Acids and Vanilloids by Lipoxygenases and Cyclooxygenases

doi: 10.1016/j.abb.2007.04.007

Figure Lengend Snippet: Percent activity of compounds relative to arachidonate

Article Snippet: Rabbit reticulocyte 15-LO-1 (r15-LO-1) and human 5-LO (h5-LO) were purchased as cell lysates from Calbiochem (La Jolla, CA) and Cayman Chemical (Ann Arbor, MI), respectively.

Techniques: Activity Assay

Journal:

Article Title: Oxidative Metabolism of Lipoamino Acids and Vanilloids by Lipoxygenases and Cyclooxygenases

doi: 10.1016/j.abb.2007.04.007

Figure Lengend Snippet: Kinetic parameters of metabolism of arachidonate and arachidonyl amino acids

Article Snippet: Rabbit reticulocyte 15-LO-1 (r15-LO-1) and human 5-LO (h5-LO) were purchased as cell lysates from Calbiochem (La Jolla, CA) and Cayman Chemical (Ann Arbor, MI), respectively.

Techniques:

Journal:

Article Title: Oxidative Metabolism of Lipoamino Acids and Vanilloids by Lipoxygenases and Cyclooxygenases

doi: 10.1016/j.abb.2007.04.007

Figure Lengend Snippet: Representative collision-induced dissociation mass spectra of NAGly and NAGABA metabolites of LO oxygenation. Reaction conditions are outlined in “Materials and Methods.” Chemical structures indicate the proposed assignments for the most abundant [M + 107Ag]+ ions. Contents of panels are as follows: A) pl12-LO with NAGly, B) pl12-LO with NAGABA, C) r15-LO-1 with NAGly, D) r15-LO-1 with NAGABA, E) h15-LO-2 with NAGly, and F) h15-LO-2 with NAGABA.

Article Snippet: Rabbit reticulocyte 15-LO-1 (r15-LO-1) and human 5-LO (h5-LO) were purchased as cell lysates from Calbiochem (La Jolla, CA) and Cayman Chemical (Ann Arbor, MI), respectively.

Techniques: