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: After washes with cold PBS and Annexin V Binding Buffer (100 mmol/l Hepes, 1.4 mol/l NaCl, 25 mmol/l CaCl 2 ), cells were resuspended in 100 μl of Annexin V Binding Buffer and incubated with 2.5 μl of PE Annexin V (BioLegend #640912) and 7 μl of 7-AAD (BD Pharmingen # 51-2359KC) for 15 min at RT in the dark.

Techniques: Cell Culture, MTT Assay, Double Staining