Journal: International Journal of Molecular Sciences

Article Title: Extracorporeal Shock Wave Therapy Alters the Expression of Fibrosis-Related Molecules in Fibroblast Derived from Human Hypertrophic Scar

doi: 10.3390/ijms19010124

Figure Lengend Snippet: The characteristics of fibroblasts derived from scar tissue (HTSFs). Matched human normal fibroblasts (HNF) and HTSF were cultured from four patients with post burn hypertrophic scar tissue. Protein expression of transforming growth factor beta 1 (TGF-β1), alpha smooth muscle actin (α-SMA), COL-Ι (collagen type Ι), COL-III (collagen type III), FN (fibronectin), Vimentin, fibroblast specific protein 1 (FSP-1), Twist-1 and N-cad (N-cadherin) was significantly higher in HTSFs compared with HNF from skin dermis. The protein expression of E-cad (E-cadherin), inhibitor of DNA binding protein 1 (ID-1) and inhibitor of DNA binding protein 2 (ID-2) were lower in HTSFs when compared with HNF from skin dermis. That expression of those proteins was measured by western blotting against specific antibody. The intensity of band was normalized with that of loading control, β-actin or lamin B1, respectively; HNF, Human normal skin derived fibroblast; HTSF, human hypertrophic scar derived fibroblast. * p < 0.05 vs. the corresponding HNF.

Article Snippet: The membranes were blocked with 5% bovine serum albumin (BSA) for 1 h at room temperature and then incubated for 16 h with polyclonal rabbit anti-TGFβ1 antibody (1:500, Santa Cruz Biotechnology, CA, USA), polyclonal mouse anti-αSMA antibody (1:500, Abcam, Cambridge, UK), monoclonal rabbit anti-fibronectin (1:2000, Abcam, Cambridge, UK), polyclonal rabbit anti-collagen-I; antibody (1:100, Abcam, Cambridge, UK), monoclonal rabbit anti-collagen-III (1:2000, Abcam, Cambridge, UK), monoclonal rabbit anti-vimentin (1:1000, Abcam, Cambridge, UK), polyclonal rabbit anti-FSP-1 antibody (1:1000, Merck Millipore, Billerica, MA, USA), monoclonal anti E-cadherin (1:1000, Cell Signaling, Danvers, MA, USA), monoclonal mouse anti N-cadherin (1:1000, Thermo Fisher Scientific, Waltham, MA, USA), monoclonal mouse anti-ID1 antibody (1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA), monoclonal mouse anti-ID2 antibody (1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and anti-β-actin (1:5000, Cell Signaling, Danvers, MA, USA).

Techniques: Derivative Assay, Cell Culture, Expressing, Binding Assay, Western Blot

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Vimentin is required for lung adenocarcinoma metastasis via heterotypic tumor cell-cancer-associated fibroblast interactions during collective invasion

doi: 10.1158/1078-0432.CCR-17-1776

Figure Lengend Snippet: (A) Immunofluorescence of vimentin (488) and FSP1 (555) on primary tumors KLV+/+ mice (scale=50μm) (B) Colocalization of vimentin and FSP1 staining (scale=10μm). (C) Quantification of percent of mice of each genotype with CAFs stratified by metastatic or non-metastatic phenotypes. (D) Immunohistochemical staining of vimentin in the metastatic and non-metastatic lymph nodes (scale bars 10× =100μm, 40× =20μm)

Article Snippet: Primary antibodies were incubated at 4°C overnight in 1% BSA in PBS (FSP1 ab27957 1:200; E-cadherin BD Biosciences 610181 1:1000).

Techniques: Immunofluorescence, Staining, Immunohistochemical staining

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Vimentin is required for lung adenocarcinoma metastasis via heterotypic tumor cell-cancer-associated fibroblast interactions during collective invasion

doi: 10.1158/1078-0432.CCR-17-1776

Figure Lengend Snippet: (A) Western blot demonstrating generation of shVIM CAFs with two different shRNA clones. (B) Immunofluorescence of vimentin in pLKO.1 andshVIM19 CAFs (scale=50μm). (C) Immunofluorescence of actin and vimentin in pLKO.1 and shVIM CAFs demonstrating cell size (10× scale= 100μm, 20× scale = 100μm). (D) Quantification of cell area of pLKO.1 and shVIM CAFs. Significance determined by unpaired t-test (p<0.05). (E) Representative images of spheroid invasion assay with pLKO.1 and shVIM CAFs in matrigel (scale = 100μm). (F) Quantification of invasive area in spheroid invasion assay. Significance determined by unpaired t-test (p<0.05). (G) Representative images ofcoculture spheroid invasion assay with H460 lung cancer line alone or in combination with pLKO.1 or shVIMCAFs (scale = 100μm). (H) Circularity of invasive areas in representative trial (3 total trials) of coculture spheroids with pLKO.1(n=6) or shVIM (n=3) CAFs. Value of 1= perfect circle. Significance determined by unpaired t-test (p<0.05). (I) Quantification of invasive chains in representative coculture spheroids with pLKO.1 or shVIM CAFs. Significance determined by unpaired t-test (p<0.05). (J) Representative images of immunofluorescence of FSP1and phalloidin in coculture spheroids. Yellow arrows mark CAFs positive for cytoplasmic FSP1 leading invasive chain of H460s (scale = 100μm).

Article Snippet: Primary antibodies were incubated at 4°C overnight in 1% BSA in PBS (FSP1 ab27957 1:200; E-cadherin BD Biosciences 610181 1:1000).

Techniques: Western Blot, shRNA, Clone Assay, Immunofluorescence, Invasion Assay

Journal: Discover. Oncology

Article Title: Cyclovirobuxine D inhibits hepatocellular carcinoma growth by inducing ferroptosis of hepatocellular carcinoma cells

doi: 10.1007/s12672-024-00940-2

Figure Lengend Snippet: CVB-D induced Ferroptosis in HepG2 and Huh-7 cells. HepG2 and Huh-7 cells were exposed to different doses of CVB-D (60, 80 and 100 µM) for 48 h. A The level of ROS was detected by fluorescent probe DCFH-DA. B The level of MDA was detected by lipid peroxidation MDA assay kit. C The protein expression of GPX4 and FSP1 was detected by western blotting. The amount of protein in term of the band intensity was analyzed by ImageJ. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs. control group

Article Snippet: The primary antibodies were diluted into 5% nonfat milk as follows: Bax antibody (ab32503, Abcam, UK, 1:5000), Bcl-2 antibody (WL01556, Wanleibio, China, 1:500), GPX4 antibody (ab125066, Abcam, UK, 1:1000), FSP1 antibody (342551, Zenbio, China, 1:1000), and GAPDH antibody (ab137959, Absin, China, 1:5000).

Techniques: Multiple Displacement Amplification, Expressing, Western Blot, Control

Journal: Discover. Oncology

Article Title: Cyclovirobuxine D inhibits hepatocellular carcinoma growth by inducing ferroptosis of hepatocellular carcinoma cells

doi: 10.1007/s12672-024-00940-2

Figure Lengend Snippet: Inhibition of ferroptosis by Fer-1 attenuates cell death induced by CVB-D in HCC cells. HepG2 and Huh-7 cells were treated with or without 100 µM CVB-D and 2 µM Fer-1 for 48 h. A The cell viability was measured by MTT assay. B The ferrous ion content was detected by a ferrous ion content assay kit. C The level of ROS was detected by fluorescent probe DCFH-DA. D The level of MDA was detected by lipid peroxidation MDA assay kit. E The protein expression of GPX4 and FSP1 was detected by western blotting. The amount of protein in term of the band intensity was analyzed by ImageJ. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs. CVB-D group

Article Snippet: The primary antibodies were diluted into 5% nonfat milk as follows: Bax antibody (ab32503, Abcam, UK, 1:5000), Bcl-2 antibody (WL01556, Wanleibio, China, 1:500), GPX4 antibody (ab125066, Abcam, UK, 1:1000), FSP1 antibody (342551, Zenbio, China, 1:1000), and GAPDH antibody (ab137959, Absin, China, 1:5000).

Techniques: Inhibition, MTT Assay, Multiple Displacement Amplification, Expressing, Western Blot

Journal: PLoS ONE

Article Title: Cadherin-9 Is a Novel Cell Surface Marker for the Heterogeneous Pool of Renal Fibroblasts

doi: 10.1371/journal.pone.0000657

Figure Lengend Snippet: Sources of the used antibodies

Article Snippet: anti FSP1/S100A4 Ab-8 , Dianova (Hamburg, Germany).

Techniques: Transduction

Journal: PLoS ONE

Article Title: Cadherin-9 Is a Novel Cell Surface Marker for the Heterogeneous Pool of Renal Fibroblasts

doi: 10.1371/journal.pone.0000657

Figure Lengend Snippet: (A) Immunohistochemical staining of two consecutive kidney cryostat sections with antisera against cadherin-9 and FSP1, a well-known renal fibroblast marker, showed that cadherin-9 positive cells also express FSP1. Staining of FSP1, however, could be also observed in the cadherin-9 negative glomerulum and in some tubular cells. The inserts in the left panel are enlarged in the right panel. (B) Cadherin-9 is not expressed by endothelial cells, as demonstrated by double immunofluorescence staining with the endothelial cell marker CD31. But cadherin-9 expression partially overlaps with CD45 as well as with α-smooth muscle actin. Cells which express only α-smooth muscle actin or CD45 are marked with an arrowhead, whereas cells which express only cadherin-9 with an arrow. Co-localization of α-smooth muscle actin or CD45 with cadherin-9 are indicated by double arrows. The different magnifications in (A) and (B) are shown by different bars.

Article Snippet: anti FSP1/S100A4 Ab-8 , Dianova (Hamburg, Germany).

Techniques: Immunohistochemical staining, Staining, Marker, Double Immunofluorescence Staining, Expressing

Journal: PLoS ONE

Article Title: Cadherin-9 Is a Novel Cell Surface Marker for the Heterogeneous Pool of Renal Fibroblasts

doi: 10.1371/journal.pone.0000657

Figure Lengend Snippet: Together with FSP1, α-smooth muscle actin (α sm actin) and CD45, cadherin-9 is a novel cell surface marker, which can be used to differentiate the depicted renal fibroblast subpopulations from other renal cell types such as epithelial and endothelial cells, smooth muscle cells, monocytes or macrophages.

Article Snippet: anti FSP1/S100A4 Ab-8 , Dianova (Hamburg, Germany).

Techniques: Marker