Journal: Molecular Medicine

Article Title: Mechanical Ventilation Augments Poly(I:C)-Induced Lung Injury via a WISP1-Integrin β3-Dependent Pathway in Mice

doi: 10.2119/molmed.2015.00233

Figure Lengend Snippet: Alterations in pulmonary levels of wisp1 mRNA expressed as (A) the ratio to the housekeeping gene β-actin and protein and (B) the ratio to the housekeeping protein β-actin at 4 h after poly(I:C) and MTV treatment. Data are presented as means ± SEM (n = 15) and compared by one-way ANOVA and Student-Newman-Keuls test. For comparing the poly(I:C) group with the untreated control group ##P < 0.01. For comparing the poly(I:C) + MTV group with the poly(I:C) group *P < 0.05, **P < 0.01.

Article Snippet: For the WISP1-blockade study, 20 mice were randomized to WISP1 antibody group or IgG group, and received 5 mg/kg body weight of anti-WISP1 neutralizing antibody (MAB1680, R&D Systems) or nonspecific isotype-specific antibody IgG (MAB002, R&D Systems) i.p., respectively.

Techniques: Control

Journal: Molecular Medicine

Article Title: Mechanical Ventilation Augments Poly(I:C)-Induced Lung Injury via a WISP1-Integrin β3-Dependent Pathway in Mice

doi: 10.2119/molmed.2015.00233

Figure Lengend Snippet: Effects of anti-WISP1 treatment on (A) EBA permeability, (B) wet:dry ratio of lungs, and (C) levels of total protein, (D) TNF-α and (E) IL-6, as well as (F) PMNs numbers in BALF at 4 h after poly(I:C) and MTV treatment. Animals received the anti-WISP1 blocking antibody or control IgG i.p. 1 h before mechanical ventilation. Data are presented as mean ± SEM (n = 10) and compared by paired t test. For comparing the anti-WISP1 group with the IgG group *P < 0.05, **P < 0.01.

Article Snippet: For the WISP1-blockade study, 20 mice were randomized to WISP1 antibody group or IgG group, and received 5 mg/kg body weight of anti-WISP1 neutralizing antibody (MAB1680, R&D Systems) or nonspecific isotype-specific antibody IgG (MAB002, R&D Systems) i.p., respectively.

Techniques: Permeability, Blocking Assay, Control

Journal: Molecular Medicine

Article Title: Mechanical Ventilation Augments Poly(I:C)-Induced Lung Injury via a WISP1-Integrin β3-Dependent Pathway in Mice

doi: 10.2119/molmed.2015.00233

Figure Lengend Snippet: Effects of poly(I:C) + MTV on WISP1/integrin β3 interaction. Lung tissues were harvested from C57BL/6J mice in the control and poly(I:C) + MTV groups. Total lysates were immunoprecipitated with an anti-WISP1 antibody or an anti-β3 antibody before immunoblot was performed. (A) Glycosylated form of integrin β3 (110 kD) was detected with an anti-integrin β3 antibody (IB: integrin β3) either in the whole lung tissue lysates (Total Lysates) or after immunoprecipitation with an anti-WISP1 antibody (IP: WISP1). (B) WISP1 was detected with an anti-WISP1 antibody (IB: WISP1) either in the whole lung tissue lysates (Total Lysates) or after immunoprecipitation with an anti-integrin β3 antibody (IP: integrin β3). Antibody species matched serum (Goat IgG) was used as a negative control for coprecipitation experiments. WISP1 and integrin β3 interact was enhanced in the poly(I:C) + MTV group as compared with the control group.

Article Snippet: For the WISP1-blockade study, 20 mice were randomized to WISP1 antibody group or IgG group, and received 5 mg/kg body weight of anti-WISP1 neutralizing antibody (MAB1680, R&D Systems) or nonspecific isotype-specific antibody IgG (MAB002, R&D Systems) i.p., respectively.

Techniques: Control, Immunoprecipitation, Western Blot, Negative Control

Journal: Molecular Medicine

Article Title: Mechanical Ventilation Augments Poly(I:C)-Induced Lung Injury via a WISP1-Integrin β3-Dependent Pathway in Mice

doi: 10.2119/molmed.2015.00233

Figure Lengend Snippet: Effects of WISP1 on poly(I:C)-induced TNF-α production in macrophages. (A) Peritoneal macrophage isolated from C57BL/6J mice were treated with WISP1 (10 μg/mL) in the presence or absence of various concentrations (0–10 μg/mL) of poly(I:C) in serum-free medium for 16 h. TNF-α levels in the supernatant were measured by ELISA. (B) Peritoneal macrophage isolated from C57BL/6J mice were treated with WISP1(10 μg/mL) in the presence or absence of poly(I:C) (5 μg/mL) in serum-free medium for up to 24 h. TNF-α levels in the supernatant were measured by ELISA at 0 h, 4 h, 8 h, 12 h, 16 h, 20 h and 24 h after stimulation. Data are presented as mean ± SEM (n = 5) and compared by two-way ANOVA. For comparing the poly(I:C) + WISP1 group with the poly(I:C) alone group *P < 0.05, ***P < 0.001.

Article Snippet: For the WISP1-blockade study, 20 mice were randomized to WISP1 antibody group or IgG group, and received 5 mg/kg body weight of anti-WISP1 neutralizing antibody (MAB1680, R&D Systems) or nonspecific isotype-specific antibody IgG (MAB002, R&D Systems) i.p., respectively.

Techniques: Isolation, Enzyme-linked Immunosorbent Assay

Journal: Molecular Medicine

Article Title: Mechanical Ventilation Augments Poly(I:C)-Induced Lung Injury via a WISP1-Integrin β3-Dependent Pathway in Mice

doi: 10.2119/molmed.2015.00233

Figure Lengend Snippet: Effects of integrin β3 knockout on WISP1 plus poly(I:C)-induced TNF-α production in macrophages. Peritoneal macrophages isolated from C57BL/6J and integrin β3 knockout mice were treated with WISP1 (10 μg/mL) and poly(I:C) (5 μg/mL) for 16 h in serum free medium. TNF-α levels in the supernatant were measured by ELISA. Data are presented as mean ± SEM (n = 5) and compared by two-way ANOVA. ***P < 0.001 C57BL/6J versus integrin β3 knockout after WISP1 plus poly(I:C) treatment.

Article Snippet: For the WISP1-blockade study, 20 mice were randomized to WISP1 antibody group or IgG group, and received 5 mg/kg body weight of anti-WISP1 neutralizing antibody (MAB1680, R&D Systems) or nonspecific isotype-specific antibody IgG (MAB002, R&D Systems) i.p., respectively.

Techniques: Knock-Out, Isolation, Enzyme-linked Immunosorbent Assay

Journal: Molecular Medicine

Article Title: Mechanical Ventilation Augments Poly(I:C)-Induced Lung Injury via a WISP1-Integrin β3-Dependent Pathway in Mice

doi: 10.2119/molmed.2015.00233

Figure Lengend Snippet: Effects of WISP1 and poly(I:C) on ERK phosphorylation in macrophages (A) Peritoneal macrophages isolated from C57BL/6J mice were treated with WISP1 (10 μg/mL) or poly(I:C) in serum free medium for 15, 30 and 60 min. ERK phosphorylation was detected by Western blot. Effects of an ERK inhibitor, U0126, on WISP1 and poly(I:C)-induced TNF-α release in macrophages. (B) Peritoneal macrophage isolated from C57BL/6J mice were treated with WISP1 (10 μg/mL) and poly(I:C) (5 μg/mL) in the presence or absence of various concentrations (0–1.0 μmol/L) of U0126 in serum free medium for 16 h. TNF-α levels in the supernatant were measured by ELISA. Data are presented as means ± SEM (n = 5) and compared by two-way ANOVA. For comparing the poly(I:C) + WISP1 group pretreated with U0126 0.5 μmol/L with poly(I:C) + WISP1 untreated with U0126 group ***P < 0.001.

Article Snippet: For the WISP1-blockade study, 20 mice were randomized to WISP1 antibody group or IgG group, and received 5 mg/kg body weight of anti-WISP1 neutralizing antibody (MAB1680, R&D Systems) or nonspecific isotype-specific antibody IgG (MAB002, R&D Systems) i.p., respectively.

Techniques: Phospho-proteomics, Isolation, Western Blot, Enzyme-linked Immunosorbent Assay