Journal: International Journal of Oncology

Article Title: Complement regulatory protein CD46 promotes bladder cancer metastasis through activation of MMP9

doi: 10.3892/ijo.2024.5659

Figure Lengend Snippet: Expression of MMP9 by CD46 in bladder cancer cells. (A) Observation of MMP2 and MMP9 expression in five bladder cancer cell lines with CD46 overexpression, analyzed by western blotting. (B) Assessment of CD46 overexpression's impact on cell growth using an in vitro proliferation assay in cells overexpressing MMP9 due to CD46. Data are presented as mean ± SD. P-values were obtained using the U Mann Whitney test ( ns P>0.05). MMP9, matrix metalloproteinase 9; veh, vehicle; MMP2, matrix metalloproteinase 2.

Article Snippet: The primary antibodies used included: CD46 (1:2,000; cat. no. ab108307; Abcam); MMP9 (1:1,000; cat. no. 7-11C; Santa Cruz Biotechnology, Inc.), phosphorylated (p-)AKT (1:1,000; cat. no. 05-669; MilliporeSigma), p38α (1:1,000; cat. no. sc-535; Santa Cruz Biotechnology, Inc.), JNK1/3 (1:1,000; cat. no. sc-474; Santa Cruz Biotechnology, Inc.), p-JNK (1:1,000; cat. no. sc-6254 Santa Cruz Biotechnology, Inc.), ERK1 (1:1,000; cat. no. sc-271269 Santa Cruz Biotechnology, Inc.), c-Fos (1:1,000; cat. no. sc-166940; Santa Cruz Biotechnology, Inc.), p-c-Fos (1:1,000; cat. no. sc-81485; Santa Cruz Biotechnology, Inc.), c-Jun (1:1,000; cat. no. sc-1694; Santa Cruz Biotechnology, Inc.), p-c-Jun (1:1,000; cat. no. sc-822; Santa Cruz Biotechnology, Inc.); p-p38 (1:2,000; cat. no. 05-1059; MilliporeSigma), AKT (1:2,000; cat. no. 9272; Cell Signaling Technology, Inc.); MMP2 (1:2,000; cat. no. NB200-114; Novus Biologicals; Bio-Techne); p-ERK (1:2,000; cat. no. 05-797R; MilliporeSigma); and β-actin (1:2,000; cat. no. 26628-22-8; MilliporeSigma).

Techniques: Expressing, Over Expression, Western Blot, In Vitro, Proliferation Assay, MANN-WHITNEY

Journal: International Journal of Oncology

Article Title: Complement regulatory protein CD46 promotes bladder cancer metastasis through activation of MMP9

doi: 10.3892/ijo.2024.5659

Figure Lengend Snippet: Effects of CD46 on the regulation of migratory and invasive potential in bladder cancer cells. (A) Media from specified cells were collected for analysis. Non-reducing gelatin SDS-PAGE was used to demonstrate gelatinase activity (top panel) and western blot analysis was conducted to show secreted CD46 (lower panel). (B) Media from cells were analyzed by ELISA assay to show secreted MMP9. (C) Bright-field images showing scratched areas (marked by white lines) in confluent monolayers of cancer cells overexpressing CD46 at different time points (left panel). Relative wound closure was assessed by measuring the width of the wounds compared with control cells (right panel). (D) Transwell assays to measure cell migration and invasion in 5637 and J82 cells transfected with CD46 (left panel). Quantification of cell migration and invasion was conducted (right panel). Each bar represents mean ± SD. All scale bars, 200 µ m. Differences between groups were assessed using a two-tailed unpaired Student's t-test. * P<0.01, ** P<0.001, *** P<0.0001 vs. control cells. veh, vehicle; MMP9, matrix metalloproteinase 9; MMP2, matrix metalloproteinase 2.

Article Snippet: The primary antibodies used included: CD46 (1:2,000; cat. no. ab108307; Abcam); MMP9 (1:1,000; cat. no. 7-11C; Santa Cruz Biotechnology, Inc.), phosphorylated (p-)AKT (1:1,000; cat. no. 05-669; MilliporeSigma), p38α (1:1,000; cat. no. sc-535; Santa Cruz Biotechnology, Inc.), JNK1/3 (1:1,000; cat. no. sc-474; Santa Cruz Biotechnology, Inc.), p-JNK (1:1,000; cat. no. sc-6254 Santa Cruz Biotechnology, Inc.), ERK1 (1:1,000; cat. no. sc-271269 Santa Cruz Biotechnology, Inc.), c-Fos (1:1,000; cat. no. sc-166940; Santa Cruz Biotechnology, Inc.), p-c-Fos (1:1,000; cat. no. sc-81485; Santa Cruz Biotechnology, Inc.), c-Jun (1:1,000; cat. no. sc-1694; Santa Cruz Biotechnology, Inc.), p-c-Jun (1:1,000; cat. no. sc-822; Santa Cruz Biotechnology, Inc.); p-p38 (1:2,000; cat. no. 05-1059; MilliporeSigma), AKT (1:2,000; cat. no. 9272; Cell Signaling Technology, Inc.); MMP2 (1:2,000; cat. no. NB200-114; Novus Biologicals; Bio-Techne); p-ERK (1:2,000; cat. no. 05-797R; MilliporeSigma); and β-actin (1:2,000; cat. no. 26628-22-8; MilliporeSigma).

Techniques: SDS Page, Activity Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Control, Migration, Transfection, Two Tailed Test

Journal: iScience

Article Title: Lupeol synergizes with 5-fluorouracil to combat c-MET/EphA2 mediated chemoresistance in triple negative breast cancer

doi: 10.1016/j.isci.2023.108395

Figure Lengend Snippet:

Article Snippet: Mouse Anti human/mouse MMP-2 (clone 8B4) , Novus , Cat# NB200-114; RRID: AB_350369.

Techniques: Recombinant, Modification, Membrane, Transfection, In Vivo, Software