Journal: Oncotarget

Article Title: Role of MIF/CD74 signaling pathway in the development of pleural mesothelioma

doi: 10.18632/oncotarget.7314

Figure Lengend Snippet: Mesothelioma cell lines studied

Article Snippet: The non-tumorigenic immortalized mesothelial cell line MeT5A and the MPM cell lines H28 (NCI-H-28), H2052 (NCI-H2052), MSTO (MSTO211H) and H2452 (NCI-H2452) were purchased from American Type Culture Collection.

Techniques:

Journal: Oncotarget

Article Title: Role of MIF/CD74 signaling pathway in the development of pleural mesothelioma

doi: 10.18632/oncotarget.7314

Figure Lengend Snippet: MIF-receptors CXCR4 mRNA ( A ), total protein ( B ) and cell surface protein ( C ) were measured in human immortalized normal mesothelial cell line (MeT5A) and different mesothelioma cell lines (JL-1, DM-3, H28, H2052, H2452, MSTO). Relative mRNA (A) or protein (B) expression levels were measured by qPCR and western blotting respectively. Data represent the mean values (± SD) of 3 to 6 independent experiments. CXCR4 distribution on the cell surface was analysed by flow cytometry (C). Cells were treated with EDTA and stained with anti-CXCR4 antibody followed by Alexa488-conjugated anti-IgG. Controls received equivalent concentrations of isotype-matched IgG. Viable mesothelial and mesothelioma cells were first gated according to SSC-A vs FSC-A scatted plot and doublet were excluded using a pulse geometry gate FSC-H × FSC-A plot (C, left panel). For all histograms, data are shown as cell number vs. the relative fluorescence. The light-coloured histogram depicts isotype control, whereas the dark-coloured one represents CXCR4 antibody. Each histogram shows data from a single representative experiment although each analysis was repeated at least seven times. CXCR4 expression was normalized according to the median fluorescence intensity with the isotype-matched IgG (C, right panels). Kruskal-Wallis test between normal mesothelial cell line MeT5A and mesothelioma cell lines: * P < 0.05.

Article Snippet: The non-tumorigenic immortalized mesothelial cell line MeT5A and the MPM cell lines H28 (NCI-H-28), H2052 (NCI-H2052), MSTO (MSTO211H) and H2452 (NCI-H2452) were purchased from American Type Culture Collection.

Techniques: Expressing, Western Blot, Flow Cytometry, Staining, Fluorescence, Control

Journal: Oncotarget

Article Title: Role of MIF/CD74 signaling pathway in the development of pleural mesothelioma

doi: 10.18632/oncotarget.7314

Figure Lengend Snippet: MIF mRNA ( A ) total protein ( B ) and secretion levels ( C ) were measured in human immortalized normal mesothelial cell line (MeT5A) and different mesothelioma cell lines (JL-1, DM-3, H28, H2052, H2452, MSTO). Relative mRNA (A) or total protein (B) expression levels were measured by qPCR and western blotting respectively. MIF concentrations in 48 h-cultured media were measured by ELISA (C). Data represent the mean values (± SD) of 3 to 6 independent experiments. Kruskal-Wallis test between normal mesothelial cell line MeT5A and mesothelioma cell lines: * P < 0.05.

Article Snippet: The non-tumorigenic immortalized mesothelial cell line MeT5A and the MPM cell lines H28 (NCI-H-28), H2052 (NCI-H2052), MSTO (MSTO211H) and H2452 (NCI-H2452) were purchased from American Type Culture Collection.

Techniques: Expressing, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Oncotarget

Article Title: Role of MIF/CD74 signaling pathway in the development of pleural mesothelioma

doi: 10.18632/oncotarget.7314

Figure Lengend Snippet: MIF-receptors CD74 mRNA ( A ), and total protein ( B ) were measured in human immortalized normal mesothelial cell line (MeT5A) and different mesothelioma cell lines (JL-1, DM-3, H28, H2052, H2452, MSTO). Relative mRNA (A) or protein (B) expression levels were measured by qPCR and western blotting respectively. Data represent the mean values (± SD) of 3 to 6 independent experiments. significant difference Kruskal-Wallis test between normal mesothelial cell line MeT5A and mesothelioma cell lines: * P < 0.05.

Article Snippet: The non-tumorigenic immortalized mesothelial cell line MeT5A and the MPM cell lines H28 (NCI-H-28), H2052 (NCI-H2052), MSTO (MSTO211H) and H2452 (NCI-H2452) were purchased from American Type Culture Collection.

Techniques: Expressing, Western Blot

Journal: Oncotarget

Article Title: Role of MIF/CD74 signaling pathway in the development of pleural mesothelioma

doi: 10.18632/oncotarget.7314

Figure Lengend Snippet: JL-1, H28 and MeT5A cells were cultured for 24 h. Cell multiplication ( A ) was measured using a MTT assay. Cell proliferation ( B ) was estimated by EdU incorporation after 2 h cultured with EdU. Proliferation rate was calculated as the percentage of EdU + -Hoechst + double-positive cells (Zeiss Apotome, 20× magnification, Axiovision 4.6). Cell apoptosis ( C ) was analyzed using Annexin-V-PE/7-AAD double staining. Effect of cisplatin on cell multiplication ( D ) was assessed using the MTT assay performed on mesothelioma cells cultured for 24 h with 100 mmol/L of cisplatin. Control was cells cultured for 24 h without cisplatin. Bars are mean values (± SD) for n = 4–15 independent experiments. Kruskal-Wallis test between normal mesothelial cell line MeT5A and mesothelioma cell lines: * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001.

Article Snippet: The non-tumorigenic immortalized mesothelial cell line MeT5A and the MPM cell lines H28 (NCI-H-28), H2052 (NCI-H2052), MSTO (MSTO211H) and H2452 (NCI-H2452) were purchased from American Type Culture Collection.

Techniques: Cell Culture, MTT Assay, Double Staining, Control

Journal: Oncotarget

Article Title: Role of MIF/CD74 signaling pathway in the development of pleural mesothelioma

doi: 10.18632/oncotarget.7314

Figure Lengend Snippet: Cell apoptosis of MeT5A, JL-1 and H28 cell lines expressed in % of total cells

Article Snippet: The non-tumorigenic immortalized mesothelial cell line MeT5A and the MPM cell lines H28 (NCI-H-28), H2052 (NCI-H2052), MSTO (MSTO211H) and H2452 (NCI-H2452) were purchased from American Type Culture Collection.

Techniques:

Journal: Oncotarget

Article Title: Role of MIF/CD74 signaling pathway in the development of pleural mesothelioma

doi: 10.18632/oncotarget.7314

Figure Lengend Snippet: H28 (pink bars) and H2052 (blue bars) cells transfected with NC-siRNA, MIF-siRNA and CD74-siRNA were assessed after 48 h of culture, for MIF expression ( A left panel), MIF secretion (A right panel) and CD74 expression ( B ) by western blot and ELISA. The multiplication of transfected H28 and H2052 cells ( C ) was evaluated by total cell counting (Hoechst-positive nuclei) after 48 h of culture. Proliferation ( D ) and apoptosis ( E ) rates of transfected H28 and H2052 cells were evaluated by EdU incorporation and TUNEL assay, respectively. Data represent the mean values (± SD) for n = 4–8 independent experiments. Kruskal-Wallis test between cells transfected with NC, MIF or CD74 siRNAs: * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: The non-tumorigenic immortalized mesothelial cell line MeT5A and the MPM cell lines H28 (NCI-H-28), H2052 (NCI-H2052), MSTO (MSTO211H) and H2452 (NCI-H2452) were purchased from American Type Culture Collection.

Techniques: Transfection, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Counting, TUNEL Assay

Journal: Oncotarget

Article Title: Role of MIF/CD74 signaling pathway in the development of pleural mesothelioma

doi: 10.18632/oncotarget.7314

Figure Lengend Snippet: Human mesothelioma cells (MSTO, JL-1, H2052 and H28) were injected s.c. (1 × 10 6 cells) and i.pl. (1 × 10 6 cells) into athymic nude mice ( n = 4–6 per group). Sub-cutaneous tumor growth rates were assessed using caliper measurements ( A ). Intra-pleural tumors were followed using [ 18 F] FDG-PET/CT. Representative PET/CT of MSTO (at 23 days post i.pl. injection) and H2052 tumors (at 69 days post i.pl. injections) in nude mice are shown ( B ). The images shown were trans-axial slices containing the FDG-avid tumors and organs, with CT (gray scale) providing anatomic references and PET (pseudo-color scale) showing the location and intensity of high tumor and organ glucose utilization. CT: CT mediastinal window; [ 18 F]FDG PET/CT: PET–CT fused image; White arrows indicated mesothelioma tumors. L = lung, H = heart, BAT = brown adipose tissue.

Article Snippet: The non-tumorigenic immortalized mesothelial cell line MeT5A and the MPM cell lines H28 (NCI-H-28), H2052 (NCI-H2052), MSTO (MSTO211H) and H2452 (NCI-H2452) were purchased from American Type Culture Collection.

Techniques: Injection, Positron Emission Tomography-Computed Tomography