Journal: PLOS One

Article Title: Pulmonary fibroblasts activated by the addition of TNF-α and IL-4 enhance lymphangiogenic capacity and ameliorate lung fibrosis in an allogeneic rat model

doi: 10.1371/journal.pone.0342528

Figure Lengend Snippet: (A) IVIS imaging performed one day after cell administration showing pulmonary localization of transplanted fibroblasts. (B) Representative Masson trichrome-stained lung sections from each group (Day 0, saline (Ctrl), rPF, and rPF + CKs). For each group, whole lung images (top row) and corresponding magnified views (bottom row) are shown to highlight differences in fibrosis and tissue architecture. (C) Quantification of fibrotic area. Data are presented as mean ± SEM. Statistical analysis was performed using the Kruskal–Wallis test, followed by Dunn’s multiple comparisons test. *p < 0.05, **p < 0.01 vs. indicated groups.

Article Snippet: Human pulmonary fibroblasts were purchased from PromoCell (Heidelberg, Germany) and cultured with HFDM-1(+) medium (Cell Science & Technology, Osaka, Japan) supplemented with 5% (v/v) Newborn Calf Serum (NBCS) and 1% (v/v) Penicillin-Streptomycin (P/S).

Techniques: Imaging, Staining, Saline

Journal: PLOS One

Article Title: Pulmonary fibroblasts activated by the addition of TNF-α and IL-4 enhance lymphangiogenic capacity and ameliorate lung fibrosis in an allogeneic rat model

doi: 10.1371/journal.pone.0342528

Figure Lengend Snippet: (A) Effect of allogeneic rPF or rPF + CKs cell treatment on plasma levels of surfactant protein D (SP-D) levels. (B) Effect of allogeneic rPF or rPF + CKs cell treatment on plasma fibrinogen levels.Pulmonary fibrosis was induced in mice (Day 0), followed by treatment with saline (Ctrl), allogeneic pulmonary fibroblasts (rPF), or cytokine-enriched fibroblasts (rPF + CKs).Plasma samples were collected on day 14 and analyzed for fibrinogen concentration using ELISA.Sample sizes (animal numbers) were as follows: SP-D: Day 0 = 8, Ctrl = 8, rPF = 8, rPF + CKs = 8 Fibrinogen: Day 0 = 7, Ctrl = 7, rPF = 7, rPF + CKs = 7.

Article Snippet: Human pulmonary fibroblasts were purchased from PromoCell (Heidelberg, Germany) and cultured with HFDM-1(+) medium (Cell Science & Technology, Osaka, Japan) supplemented with 5% (v/v) Newborn Calf Serum (NBCS) and 1% (v/v) Penicillin-Streptomycin (P/S).

Techniques: Clinical Proteomics, Saline, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Combined HIF-1α blockade and CHIR99021 treatment reverses pulmonary fibrosis via modulation endothelial-to-mesenchymal transition

doi: 10.1016/j.isci.2025.114028

Figure Lengend Snippet: Combination treatment of CHIR99021 and 2-ME reverses radiation-induced fibrotic changes, reducing persistent DNA damage in endothelial cells and fibroblasts (A) Schematic representation of irradiation and CHIR99021 treatment in HUVECs. HUVECs were treated with CHIR99021 (3 μM) after 3 days of irradiation (10 Gy). Immunofluorescence staining for phalloidin and VE-cadherin detection was performed 6 days post-irradiation (magnification: 400×). Scale bars = 20 μm. Bar graphs quantify phalloidin intensity. (B) Schematic representation of 2-ME and CHIR99021 treatment and irradiation in HUVECs. HUVECs were irradiated (10 Gy) and treated with 2-ME (0.5 μM) after 2 days, followed by CHIR99021 (3 μM) after 3 days. Immunofluorescence staining and quantification of γH2AX in HUVECs was performed 5 days post-irradiation (magnification: 400×); Scale bars = 10 μm; scale bar of cropped images = 5 μm. Dot graph quantifies the number of γH2AX foci in more than 100 cells. (C) Schematic representation of irradiation (10 Gy), 2-ME (0.5 μM), CHIR99021 (3 μM), and hrVEGF (30 ng/mL) treatment in HUVECs. Cell morphology and phalloidin immunofluorescence staining were performed 11 days post-irradiation (magnification: 200×). Scale bars = 300 μm. Bar graphs quantify phalloidin intensity. (D) Immunofluorescence staining and quantification of OCT4, Sox2, and Nanog in HUVECs 11 days post-irradiation (magnification: 400×). Scale bars = 5 μm. Dot graph quantifies OCT4, Sox2, and Nanog intensity in more than 100 cells. (E) HUVECs, irradiated for 48 h, were cultured on Matrigel-coated plates for 3 h in the presence of CHIR99021, 2-ME, CHIR99021+2-ME, or medium only as a control (magnification: 40×). Scale bars = 20 μm. Bar graphs quantify the number of tubes formed. (F) Schematic representation of irradiation (10 Gy) and 2-ME (0.5 μM) and CHIR99021 (3 μM) treatment in HPFs. Cell morphology and phalloidin immunofluorescence staining were performed 11 days post-irradiation (magnification: 200×). Scale bars = 300 μm. Bar graphs quantify phalloidin intensity (right panels). (G) Immunofluorescence staining and quantification of γH2AX in HPFs 11 days post-irradiation (magnification: 400×). Scale bars = 5 μm. Dot graph quantifies the number of γH2AX foci in more than 100 cells. (H) Immunofluorescence staining and quantification of OCT4, Sox2, and Nanog in HPFs 11 days post-irradiation (magnification: 400×). Scale bars = 5 μm. Dot graph quantifies OCT4, Sox2, and Nanog intensity in more than 100 cells. In all graphs, error bars indicate SEM. Statistical significance was assessed by one-way ANOVA for multiple comparisons. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. In graphs (D) and (G), error bars represent SD. The data shown are representative of repeated three independent experiments.

Article Snippet: Human pulmonary fibroblasts (HPFs) , Promocell , Cat# C-12360.

Techniques: Irradiation, Immunofluorescence, Staining, Cell Culture, Control