Journal: JBMR plus

Article Title: Rapamycin does not alter bone microarchitecture or material properties quality in young-adult and aged female C57BL/6 mice.

doi: 10.1093/jbmrpl/ziae001

Figure Lengend Snippet: Figure 3. Effects of age and rapamycin on (A) serum CTX-1, (B) serum P1NP, (C) serum P1NP:CTX-1, (D and E) immunoblotting analysis and quantification of tibia OPG:RANKL, (F) “matrix-homeostasis” multi-plex protein array, (G) “inflammatory” multi-plex protein array, (H-K) immunoblotting analysis, and quantification of phosphorylated S6 (S240/244) and AKT (S473) in tibia of 5- and 22 mo-old female C57BL/6 mice following 8 wk of treatment with rapamycin (2 mg/kg; 1 × 5d) or vehicle. Data are presented as mean (white circle), median (line), 25%/75% IQR (bottom and top of box), and minimum/maximum range (whiskers), with individual values (black circles) overlayed.

Article Snippet: Antibodies used were S6 Ribosomal Protein (2217; Cell Signaling Technology; 1:1000 dilution in Tris-Buffered Saline with 0.1% Tween 20 [TBST] and 5% bovine serum albumin [BSA]), serine 240/244 phospho-S6 Ribosomal protein (2215; Cell Signaling Technology; 1:1000 dilution in TBST with 5% BSA), AKT (4691; Cell Signaling Technology; 1:1000 dilution in TBST with 5% BSA), serine 473 phospho-AKT (4060; Cell Signaling Technology; 1:2000 dilution in TBST with 5% BSA), Osteoprotegerin (OPG; AF459; R&D Systems; 1:1000 dilution in TBST with 5% nonfat milk), and RANKL (sc-377 079; Santa Cruz Biotechnology; 1:200 dilution in TBST with 5% BSA).

Techniques: Western Blot, Protein Array

Journal: Respiratory Research

Article Title: Lipoxin A 4 ameliorates lipopolysaccharide-induced lung injury through stimulating epithelial proliferation, reducing epithelial cell apoptosis and inhibits epithelial–mesenchymal transition

doi: 10.1186/s12931-019-1158-z

Figure Lengend Snippet: LXA4 stimulates AT II cell proliferation and reduces AT II cell apoptosis in LPS induced lung injury. C57BL/6 J mice were intra-tracheal given N.S or LPS 10 mg/kg for 24 h, with or without LXA4 1μg intraperitoneal injection per mouse. Immunofluorescence staining of lung specimens was shot by fluorescence microscope and calculated by positive goals compared to DAPI. a - b : co-dying of SP-C and PCNA (× 200, × 400). c - d : co-dying of SP-C and TUNEL (× 200, × 400), scar bar = 50 μm. Data were presented with means ± SD; n = 4; *** P < 0.001, **** P < 0.0001

Article Snippet: HAT II cells were pre-incubated with or without BOC-2 10μΜ or BML-111 10μΜ 30 min before TGF-β 1 10 ng/ml for 48 h with or without LXA4 100 nM. a - d : the effect of LXA4 on EMT was promoted by the pre-incubation of AT II cells with BML-111 (the LXA4 receptor agonist). e - h : the effect of LXA4 on EMT was abrogated by the pre-incubation of AT II cells with BOC-2(the LXA4 receptor antagonist). n = 4 for each culture condition, repeated using cells from 4 donors.

Techniques: Injection, Immunofluorescence, Staining, Fluorescence, Microscopy, TUNEL Assay

Journal: Respiratory Research

Article Title: Lipoxin A 4 ameliorates lipopolysaccharide-induced lung injury through stimulating epithelial proliferation, reducing epithelial cell apoptosis and inhibits epithelial–mesenchymal transition

doi: 10.1186/s12931-019-1158-z

Figure Lengend Snippet: LXA4 alleviates apoptosis in LPS-induced lung injury. C57BL/6 J mice were intra-tracheal given N.S or LPS 10 mg/kg for 24 h, with or without LXA4 1μg per mouse intraperitoneal injection. Immunofluorescence staining of lung specimens was shot by fluorescence microscope and calculated by positive goals compared to DAPI. a - b : Immunofluorescence staining of cleaved-caspase-3, scar bar = 50 μm. c - d : WB of cleaved-caspase-3. Data were presented with means ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001; n = 3

Article Snippet: HAT II cells were pre-incubated with or without BOC-2 10μΜ or BML-111 10μΜ 30 min before TGF-β 1 10 ng/ml for 48 h with or without LXA4 100 nM. a - d : the effect of LXA4 on EMT was promoted by the pre-incubation of AT II cells with BML-111 (the LXA4 receptor agonist). e - h : the effect of LXA4 on EMT was abrogated by the pre-incubation of AT II cells with BOC-2(the LXA4 receptor antagonist). n = 4 for each culture condition, repeated using cells from 4 donors.

Techniques: Injection, Immunofluorescence, Staining, Fluorescence, Microscopy

Journal: Respiratory Research

Article Title: Lipoxin A 4 ameliorates lipopolysaccharide-induced lung injury through stimulating epithelial proliferation, reducing epithelial cell apoptosis and inhibits epithelial–mesenchymal transition

doi: 10.1186/s12931-019-1158-z

Figure Lengend Snippet: LXA4 stimulates HAT II cell proliferation and reduces HAT II cell apoptosis in vitro. HAT II cells were cultured as mentioned in the methods. a and b : apoptosis of HAT II after the stimulation of LPS and LXA4. c : Proliferation of HAT II after the stimulation of LPS and LXA4. Data were presented with means ±SEM. ** P < 0.01, *** P < 0.001, **** P < 0.0001, n = 4 for each culture condition, repeated using cells from 4 donors

Article Snippet: HAT II cells were pre-incubated with or without BOC-2 10μΜ or BML-111 10μΜ 30 min before TGF-β 1 10 ng/ml for 48 h with or without LXA4 100 nM. a - d : the effect of LXA4 on EMT was promoted by the pre-incubation of AT II cells with BML-111 (the LXA4 receptor agonist). e - h : the effect of LXA4 on EMT was abrogated by the pre-incubation of AT II cells with BOC-2(the LXA4 receptor antagonist). n = 4 for each culture condition, repeated using cells from 4 donors.

Techniques: In Vitro, Cell Culture

Journal: Respiratory Research

Article Title: Lipoxin A 4 ameliorates lipopolysaccharide-induced lung injury through stimulating epithelial proliferation, reducing epithelial cell apoptosis and inhibits epithelial–mesenchymal transition

doi: 10.1186/s12931-019-1158-z

Figure Lengend Snippet: LXA4 inhibits the epithelial-mesenchymal transition (EMT) in LPS-induced lung injury. C57BL/6 J mice were intra-tracheal given N.S or LPS 10 mg/kg (for 24 h, 48 h or 72 h), with or without LXA4 1μg per mouse intraperitoneal injection. Immunofluorescence staining of lung specimens was shot by fluorescence microscope and calculated by positive goals compared to DAPI. a - h : Immunofluorescence staining of marker of EMT (× 400): E-cadherin( a - b ), N-cadherin( c - d ), α-SMA( e - f ) and Vimentin( g - h ). i and j : co-dying of SP-C and α-SMA. scar bar = 50 μm. All of the data was conducted in triplicate. Data were presented with means ± SD. n = 3. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: HAT II cells were pre-incubated with or without BOC-2 10μΜ or BML-111 10μΜ 30 min before TGF-β 1 10 ng/ml for 48 h with or without LXA4 100 nM. a - d : the effect of LXA4 on EMT was promoted by the pre-incubation of AT II cells with BML-111 (the LXA4 receptor agonist). e - h : the effect of LXA4 on EMT was abrogated by the pre-incubation of AT II cells with BOC-2(the LXA4 receptor antagonist). n = 4 for each culture condition, repeated using cells from 4 donors.

Techniques: Injection, Immunofluorescence, Staining, Fluorescence, Microscopy, Marker

Journal: Respiratory Research

Article Title: Lipoxin A 4 ameliorates lipopolysaccharide-induced lung injury through stimulating epithelial proliferation, reducing epithelial cell apoptosis and inhibits epithelial–mesenchymal transition

doi: 10.1186/s12931-019-1158-z

Figure Lengend Snippet: LXA4 inhibits TGF-β 1 induced EMT in primary HAT II cells. HAT II cells were incubated with or without TGF-β 1 10 ng/ml for 48 h with or without LXA4 0.1 nM, 1 nM, 10 nM and 100 nM. a - g : the mRNA expression of CDH-1, SP-C, AQP-5, CDH-2, α-SMA and fibronectin. h - k : the protein expression level of E-cadherin, N-cadherin and α-SMA. n = 4 for each culture condition, repeated using cells from 4 donors. Data were presented with means ±SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: HAT II cells were pre-incubated with or without BOC-2 10μΜ or BML-111 10μΜ 30 min before TGF-β 1 10 ng/ml for 48 h with or without LXA4 100 nM. a - d : the effect of LXA4 on EMT was promoted by the pre-incubation of AT II cells with BML-111 (the LXA4 receptor agonist). e - h : the effect of LXA4 on EMT was abrogated by the pre-incubation of AT II cells with BOC-2(the LXA4 receptor antagonist). n = 4 for each culture condition, repeated using cells from 4 donors.

Techniques: Incubation, Expressing

Journal: Respiratory Research

Article Title: Lipoxin A 4 ameliorates lipopolysaccharide-induced lung injury through stimulating epithelial proliferation, reducing epithelial cell apoptosis and inhibits epithelial–mesenchymal transition

doi: 10.1186/s12931-019-1158-z

Figure Lengend Snippet: LXA4 inhibits TGF-β 1 -induced EMT in primary AT II cells through activation of LXA4 receptor (ALX). HAT II cells were pre-incubated with or without BOC-2 10μΜ or BML-111 10μΜ 30 min before TGF-β 1 10 ng/ml for 48 h with or without LXA4 100 nM. a - d : the effect of LXA4 on EMT was promoted by the pre-incubation of AT II cells with BML-111 (the LXA4 receptor agonist). e - h : the effect of LXA4 on EMT was abrogated by the pre-incubation of AT II cells with BOC-2(the LXA4 receptor antagonist). n = 4 for each culture condition, repeated using cells from 4 donors. Data were presented with means ±SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: HAT II cells were pre-incubated with or without BOC-2 10μΜ or BML-111 10μΜ 30 min before TGF-β 1 10 ng/ml for 48 h with or without LXA4 100 nM. a - d : the effect of LXA4 on EMT was promoted by the pre-incubation of AT II cells with BML-111 (the LXA4 receptor agonist). e - h : the effect of LXA4 on EMT was abrogated by the pre-incubation of AT II cells with BOC-2(the LXA4 receptor antagonist). n = 4 for each culture condition, repeated using cells from 4 donors.

Techniques: Activation Assay, Incubation

Journal: Respiratory Research

Article Title: Lipoxin A 4 ameliorates lipopolysaccharide-induced lung injury through stimulating epithelial proliferation, reducing epithelial cell apoptosis and inhibits epithelial–mesenchymal transition

doi: 10.1186/s12931-019-1158-z

Figure Lengend Snippet: LXA4 reduces TGF-β 1 -induced EMT in HAT II cells partly through the SMAD and PI3K/AKT signaling pathways. HAT II cells were pre-incubated with 10 μM Sis3 (a specific Smad3 inhibitor), and 10 μM LY294002(PI3Kinhibitor) for 30 min prior to TGF-β 1 10 ng/ml for 48 h with or without LXA4 100 nM. a - h : Sis3 and LY294002 treatment abolished the inhibition of LXA4 on the EMT in AT II cells. i - k : LXA4 inhibited TGF-β 1 -induced the phosphorylation of AKT and Smad in primary AT II cells. n = 4 for each culture condition, repeated using cells from 4 donors. Data were presented with means ±SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: HAT II cells were pre-incubated with or without BOC-2 10μΜ or BML-111 10μΜ 30 min before TGF-β 1 10 ng/ml for 48 h with or without LXA4 100 nM. a - d : the effect of LXA4 on EMT was promoted by the pre-incubation of AT II cells with BML-111 (the LXA4 receptor agonist). e - h : the effect of LXA4 on EMT was abrogated by the pre-incubation of AT II cells with BOC-2(the LXA4 receptor antagonist). n = 4 for each culture condition, repeated using cells from 4 donors.

Techniques: Incubation, Inhibition