Journal: Cancer

Article Title: Colorectal carcinoma cell production of transforming growth factor beta decreases expression of endothelial cell vascular endothelial growth factor receptor 2.

doi: 10.1002/cncr.26247

Figure Lengend Snippet: Figure 6. (A) Representative Western blot shows vascular endothelial growth factor receptor 2 (VEGFR2) and activated Smad protein expression and (B) proportion of VEGFR2 protein levels in vehicle-treated and 5 ng/mL transforming growth factor beta (TGF-b)-treated bovine aortic endothelial cells (BAECs) cultured in atmospheric (21%) O2 and <0.1% O2 (anoxia) over 6 and 24 hours. Analysis of variance, P ¼ .0015; n ¼ 4; *P < .05. (C) VEGFR2 and Smad protein levels in BAECs treated for 24 hours with 0 to 50 ng/mL bone morphogenetic protein 9 (BMP9) are shown. (D) VEGFR2 and Smad protein levels in BAECs treated for 24 hours with combinations of 10 ng/mL BMP9, 5 ng/mL TGF-b1, and 5 lM SB-431542 (SB) are shown. Western blot is representative of 2 independent experiments.

Article Snippet: BAECs were grown until confluent and serum-starved overnight before treatment in serum-free medium with 0.1 to 20 ng/mL of recombinant human TGF-b1; 10 ng/mL recombinant human BMP9 (both R&D Systems, Minneapolis, Minn); 5 lM SB431542, an inhibitor of TGbR-I isoforms ALK5, ALK4, and ALK722; and 2.5 lM SD-208, an inhibitor of ALK5 and ALK4 (both Tocris Bioscience, Ellisville, Mo).23 CRC cells were grown until superconfluent, serumstarved overnight, then incubated in fresh serum-free DMEM for 48 hours.

Techniques: Western Blot, Expressing, Cell Culture

Journal: British journal of cancer

Article Title: Inactivation of the WASF3 gene in prostate cancer cells leads to suppression of tumorigenicity and metastases.

doi: 10.1038/sj.bjc.6605850

Figure Lengend Snippet: Figure 7 Knockdown of WASF3 expression suppresses MMP9 expression levels and inhibits MMP9 release from prostate cancer cell lines. (A) RT–PCR analysis of PC3 and DU145 cells showing knockdown of WASF3 also show down-regulation of MMP9, but not MMP2 expression levels. Cont ¼ parental cells. (B) Secretion of the pro- and active form of MMP9 from PC3 and DU145 cells were measured in the supernatants of actively growing cells using the Fluorokine MAP assay procedure. Cells from each clone showing knockdown of WASF3 also showed reduced MMP9 levels compared with parental (control) and non-knockdown cells that did not.

Article Snippet: After 24 h, the media were collected in tubes and centrifuged for 10 min at 10 000 g. The pro- and active MMP-9 levels released into the media were measured using a Fluorokine MAP human MMP9 kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Knockdown, Expressing, Reverse Transcription Polymerase Chain Reaction, Control