Journal: American Journal of Physiology - Cell Physiology

Article Title: WISP1 induces the expression of macrophage migration inhibitory factor in human lung fibroblasts through Src kinases and EGFR-activated signaling pathways

doi: 10.1152/ajpcell.00410.2023

Figure Lengend Snippet: αvβ5-integrin and chondroitin sulfate proteoglycans are implicated in Wnt1-inducible signaling protein 1 (WISP1)-induced production of migration inhibitory factor (MIF) from human lung fibroblasts (HLFs). HLFs were pretreated for 30 minutes with anti-αvβ5-integrin antibody (6 µg/mL) ( A ) or for 2 h with chondroitinase ABC or chondroitinase AC II (3 units/mL) ( B ), followed by stimulation with WISP1 (100 ng/mL) for 48 h, and the levels of MIF in conditioned media were determined by ELISA. Data represent the means ± SD ( n = 3). *Statistically significant increase ( P < 0.05) compared to control. #Statistically significant decrease ( P < 0.05) compared to WISP1-treated cells. IgG represents immunoglobulins from nonimmunized mouse.

Article Snippet: Recombinant human EGF and anti-integrin αvβ5 mouse mAb (no. MAB2528-SP) were purchased from R&D Systems (Bio-Techne Ltd., Abingdon, UK).

Techniques: Migration, Enzyme-linked Immunosorbent Assay, Control

Journal: American Journal of Physiology - Cell Physiology

Article Title: WISP1 induces the expression of macrophage migration inhibitory factor in human lung fibroblasts through Src kinases and EGFR-activated signaling pathways

doi: 10.1152/ajpcell.00410.2023

Figure Lengend Snippet: Proposed model of the effects of Wnt1-inducible signaling protein 1 (WISP1) on the expression of migration inhibitory factor (MIF) and its receptors, as well as on the expression of cyclooxygenase (COX)-2, IL-6, and matrix metalloproteinase (MMP)-2, and production of prostaglandin E 2 (PGE 2 ) from human lung fibroblasts (HLFs). The effect of WISP-1 on the expression of MIF in HLFs with the complex interactions between WISP-1, αvβ5, Src kinases, EGF receptor (EGFR), and CD74/CD44 receptors, reveal intricate regulatory networks that could contribute to the expression of proinflamamtory molecules and remodeling enzymes secretion.

Article Snippet: Recombinant human EGF and anti-integrin αvβ5 mouse mAb (no. MAB2528-SP) were purchased from R&D Systems (Bio-Techne Ltd., Abingdon, UK).

Techniques: Expressing, Migration

Journal: Journal of Oncology

Article Title: Anoikis Resistance as a Further Trait of Acidic-Adapted Melanoma Cells

doi: 10.1155/2019/8340926

Figure Lengend Snippet: An anoikis resistant-like phenotype in acidic melanoma cells. (a) Representative images of western blot for N-cad, EGFR, pAKT/AKT, pERK/ERK, Tie1, IKB, and β -actin of A375M6 melanoma cells exposed to standard medium (pH 7.4), to an acidified medium for 24 hours (transient exposure, pH 6.7) or to a reduced pH medium, for approximately three months (chronic exposure, pH 6.7c), and (right) densitometry graph of protein expression. (b) Representative images (left) of invasiveness of melanoma cells grown in different pH conditions in the presence or absence of 25 μ M Ilomastat (a metalloproteinases inhibitor) and quantitative analysis (right) of the number of cells that migrated through Matrigel. (c) Representative images (left) of flow cytometric analysis of α v β 3 and of α v β 5 integrin expression of A375M6 melanoma cells grown in different pH conditions and (right) quantitative analysis of integrin expression as percentage of increment in Mean Fluorescent Intensity. Each experiment was conducted in triplicate and data are expressed as mean ± SEM of at least three independent experiments. ∗ p<0.05.

Article Snippet: Mouse anti-human integrin receptor α v β 3 (clone LM609, Millipore) and mouse anti-human integrin receptor α v β 5 (sc13588, Sigma), mouse monoclonal AF6-88.5.3 specific for class I H-2Kb antigen (provided by Dr. S. Gattoni-Celli, Medical University of South Carolina, USA) (Calorini 1999) or mouse monoclonal HLA-ABC antigen clone W6/32 (DAKO), were used.

Techniques: Western Blot, Expressing

Journal: Cell

Article Title: Type V collagen in scar tissue regulates the size of scar after heart injury

doi: 10.1016/j.cell.2020.06.030

Figure Lengend Snippet: (A) Expression of ECM and myofibroblast genes in Col5a1CKO CFs generated ex vivo (n=6) (B,C) Flow cytometry to determine expression of (B) αvβ3 and (C) αvβ5 integrins on Col5a1CKO CFs (n=6). (D,E) Immunostaining for Vim and (D) αvβ3, (E) αvβ5 in scar tissue at 7 days post-MI (arrows, representative images) (F) Expression of key myofibroblast genes in Col5a1CKO CFs in the presence or absence of cilengitide (n=6). (G) Experimental design to treat animals with daily cilengitide (20mg/kg) (H) EF/FS in control and Col5a1CKO injected with cilengitide or vehicle (*Col5a1CKO+Cilengitide [red dotted line] vs. Col5a1CKO+Veh [red solid line], n=13/CKO+Cilengitide 10/other groups at basal, n=12/CKo+Cilengitide, 6/CKO+Veh, 9/Control+Cilengitide, 7/Control+Veh at 2wks post MI). (I) Representative images of M mode echocardiogram (yellow line indicates end systolic diameter) (J) Masson trichrome staining of mid ventricle at 2 weeks post-MI to show scar size (arrowhead, n=same number at 2 weeks post-MI as above) (K) Quantitation of fibrotic area (n=same number as above) (L) Fraction of Col5a1CKO animals demonstrating mild, moderate and severe fibrosis following PBS or cilengitide infusion (M) Immunostaining for αSMA and Vimentin in hearts of Col5a1CKO receiving PBS or cilengitide and quantitation of the fraction (arrows, representative images, n=10/CKO+Cilengitide, n=6/CKO+Veh, n=6/animals for all other groups). (N) Dot plot representing expression of ECM genes that are upregulated in CFs of Col5a1CKO hearts at 7 days post-MI compared to controls (left panel), the same genes were shown in fibroblasts from Col5a1CKO+Vehicle and Col5a1CKO+Cilengitide samples (right panel). (O) Box plot showing the module scores of 28 genes from (N) in fibroblasts from Col5a1CKO+Veh and Col5a1CKO+Cilengitide. Data shown as mean± S.D., *p<0.05, ns: Not significant.

Article Snippet: Antibodies and probes The following primary antibodies, reagents, or probes were used for immunostaining: rabbit anti-Vimentin (1:100, Abcam, ab45939); mouse anti-smooth muscle actin (1:100, Dako, M0851); anti-cardiac Troponin I (1:100, Abcam, ab47003); mouse anti-integrin αVβ3 (1:50, Abcam, ab7166); mouse integrin αVβ5 (1:20, R&D, MAB2528); Alexa Fluor 594 conjugated WGA (5μg/ml, Invitrogen, {"type":"entrez-nucleotide","attrs":{"text":"W11262","term_id":"1285567","term_text":"W11262"}} W11262 ).

Techniques: Expressing, Generated, Ex Vivo, Flow Cytometry, Immunostaining, Injection, Staining, Quantitation Assay