Journal: Redox Biology

Article Title: Identification of tyrosine brominated extracellular matrix proteins in normal and fibrotic lung tissues

doi: 10.1016/j.redox.2024.103102

Figure Lengend Snippet: PXDN and laminin expression are modulated in human lung fibroblasts (HLF) and bone-marrow-derived macrophages (BMDM) upon TGF-β1 stimulation. (A – C) HLFs were treated 5 ng/mL TGF-β1 for 48 h, and analyzed for LAMA1 (A) or PXDN (B) mRNA or PXDN protein expression (C). (D – G) BMDM from C57BL6/NJ mice were treated 5 ng/mL TGF-β1 for 48 h, and analyzed for Pxdn (D) , Lama1 (E) , or Lamb1 (F) mRNA, or Laminin α/β1 protein expression by immunofluorescence imaging (G). Scale bar: 50 μm. Values are the mean of at least three independent biological replicates ± SEM. Differences among groups were evaluated by Student's T-test.

Article Snippet: HLF (ATCC PCS-201-013) were cultured in fibroblast basal medium (ATCC PCS-201-030) with fibroblast growth kit-low serum (ATCC PCS-201-041) at 37 °C in a 5% CO 2 atmosphere.

Techniques: Expressing, Derivative Assay, Immunofluorescence, Imaging

Journal: Scientific Reports

Article Title: Plant miRNA osa-miR172d-5p suppressed lung fibrosis by targeting Tab1

doi: 10.1038/s41598-023-29188-6

Figure Lengend Snippet: Osa-miR172d-5p identified as a plant miR candidate with an anti-fibrotic effect. ( A ) Scheme of plant miR selection. ( B ) In silico analysis of the interaction between osa-miR172d-5p and TAB1. ( C ) Human lung fibroblast HFL1 cells were transfected with the osa-miR172d-5p for 48 h, and TAB1 expression was evaluated via western blot analysis ( n = 4). ( D ) HFL1 cells were transfected with the indicated concentration of osa-miR172d-5p for 48 h, and cDNA was evaluated via qRT-PCR ( n = 4). Data are shown as mean ± SEM. * P < 0.05. *** P < 0.001 versus control group.

Article Snippet: Human lung fibroblast HFL1 cells (JCRB, Osaka, Japan) were cultured in 10% fetal bovine serum (FBS) (Sigma-Aldrich), Dulbecco's Modified Eagle Medium (DMEM) (044-29765; Fujifilm Tokyo, Japan) supplemented with penicillin G (876111)–streptomycin (876161; Meiji Pharmaceutical Co., Tokyo, Japan) in 5% CO 2 and 100% humidity at 37 °C.

Techniques: Selection, In Silico, Transfection, Expressing, Western Blot, Concentration Assay, Quantitative RT-PCR, Control

Journal: Scientific Reports

Article Title: Plant miRNA osa-miR172d-5p suppressed lung fibrosis by targeting Tab1

doi: 10.1038/s41598-023-29188-6

Figure Lengend Snippet: TAB1 knockdown suppressed TGFβ-induced fibrotic gene expression. ( A ) Human lung fibroblast HFL1 cells were transfected with TAB1-siRNA (10 nM, 48 h) and TAB1 expression levels were determined by western blot analysis. ( B – D ) Human lung fibroblast HFL1 cells were transfected with TAB1-siRNA (10 nM, 48 h) and treated with TGFβ (5 ng/mL for B , D 48 h; C 24 h). mRNA expression levels were assessed via RT-qPCR. ( B ) ASMA (αSMA) (n = 4), ( C ) COL1A1 (n = 4), and ( D ) FN (fibronectin; n = 4). Data are shown as mean ± SEM. * P < 0.05. ** P < 0.01. *** P < 0.001 versus control group.

Article Snippet: Human lung fibroblast HFL1 cells (JCRB, Osaka, Japan) were cultured in 10% fetal bovine serum (FBS) (Sigma-Aldrich), Dulbecco's Modified Eagle Medium (DMEM) (044-29765; Fujifilm Tokyo, Japan) supplemented with penicillin G (876111)–streptomycin (876161; Meiji Pharmaceutical Co., Tokyo, Japan) in 5% CO 2 and 100% humidity at 37 °C.

Techniques: Knockdown, Gene Expression, Transfection, Expressing, Western Blot, Quantitative RT-PCR, Control

Journal: Cancer Science

Article Title: Elevated expression of angiomodulin (AGM/IGFBP‐rP1) in tumor stroma and its roles in fibroblast activation

doi: 10.1111/j.1349-7006.2012.02203.x

Figure Lengend Snippet: Effect of transforming growth factor‐β1 (TGF‐β1) on expression of angiomodulin (AGM), fibronectin (FN) and α‐smooth muscle actin (α‐SMA) in two kinds of cultured human fibroblasts. Human natal dermal fibroblasts (HDFs) (a) and WI38 cells (b) were incubated with the indicated concentrations (ng/mL) of TGF‐β1 in serum‐free medium for 2 days. From each culture, the conditioned medium and cell lysates were prepared, as described in Materials and Methods. AGM and fibronectin were analyzed with the conditioned media, while α‐SMA and β‐actin as an internal loading control were done with the cell lysates. The results were reproduced in at least three separate experiments.

Article Snippet: Human fetal lung fibroblast cell line WI38 and human bladder carcinoma cell line T24 (EJ‐1 strain) were provided from Japanese Collection of Research and Bioresources (JCRB, Osaka, Japan).

Techniques: Expressing, Cell Culture, Incubation, Control

Journal: Cancer Science

Article Title: Elevated expression of angiomodulin (AGM/IGFBP‐rP1) in tumor stroma and its roles in fibroblast activation

doi: 10.1111/j.1349-7006.2012.02203.x

Figure Lengend Snippet: Effects of angiomodulin (AGM) and transforming growth factor‐β1 (TGF‐β1) on growth of human fibroblasts. (a,b) Human natal dermal fibroblasts (HDFs) were incubated with the indicated concentrations of TGF‐β1 for 5 days (a) or AGM for 4 days (β) in DMEM/F12+5% FCS medium on 24‐well plates. Each point represents the mean ± SD of the numbers of cells in triplicate wells. (c) Time course of HDF growth in presence (●) or absence (○) of 10 μg/mL AGM. (d) Effect of varied concentrations of AGM on the growth of HDFs was examined in the presence (●) or absence (○) of 10 μM Smad inhibitor SB431542 (Smad inh.) for 6 days on a 96‐well plate. The cell growth was measured by the crystal violet staining. Each point represents the mean ± SD in triplicate wells. (e,f) Effects of varied concentrations of TGF‐β1 (e) or ΑGΜ (f) on the growth of WI38 cells were examined for 5 days as described in (a) and (b). Other experimental conditions are described in Materials and Methods.

Article Snippet: Human fetal lung fibroblast cell line WI38 and human bladder carcinoma cell line T24 (EJ‐1 strain) were provided from Japanese Collection of Research and Bioresources (JCRB, Osaka, Japan).

Techniques: Incubation, Staining