primary antibodies against 15lo1  (Novus Biologicals)

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