Journal: Oncotarget

Article Title: Hypoxia promotes colon cancer dissemination through up-regulation of cell migration-inducing protein (CEMIP)

doi:

Figure Lengend Snippet: A. Western blot analysis of whole cell lysates from cells as indicated using antibodies directed against CEMIP and Jarid1A. Tubulin was used as a loading control. B & C. Real Time RT-PCR and Western blot analysis of HCT-116 (B) and MDA-MB-231 (C) cells for ectopically expressed Jarid1A and endogenous CEMIP. D. Western blot analysis was used to determine the silencing efficiency of Jarid1A shRNAs in SW480 cells (left panel). Real Time RT-PCR using the most effective Jarid1A shRNA2 was performed to monitor the effect of Jarid1A on CEMIP expression (middle panel). Transwell chamber migration assay was performed in SW480 cells expressing Jarid1A shRNA2 or control shRNA (right panel). After staining with DAPI for nuclear, the migrated cells were counted in 10 different fields using a Nikon 10x lens. E. Decreased levels of H3K4me3 within the CEMIP promoter region in MDA-MB-231 cells overexpressing Jarid1A. A ChIP assay followed by quantitative PCR was performed in MDA-MB-231 cells trasnfected with Jarid1A cDNA or vector control. An anti-H3K4me3 antibody or an IgG control antibody was used for the ChiP.

Article Snippet: HIF-1α P402A/P564A (plasmid 18955), HIF-2α P405A/P531A (plasmid 18956), and Jarid1A plasmids (plasmid 14800) were purchased from Addgene.

Techniques: Western Blot, Control, Quantitative RT-PCR, Expressing, Migration, shRNA, Staining, Real-time Polymerase Chain Reaction, Plasmid Preparation

Journal: Oncotarget

Article Title: 14-3-3ζ promotes hepatocellular carcinoma venous metastasis by modulating hypoxia-inducible factor-1α

doi: 10.18632/oncotarget.7493

Figure Lengend Snippet: ( A ) The interaction between HIF-1α and 14-3-3ζ was determined by co-IP assays in SMMC-7721 and CSQT-2 cells over-expressing 14-3-3ζ expression or treated with CoCl2. Cell-lysates was immunoprecipitated with anti-14-3-3ζ antibody, and then probed using anti-HIF-1α antibody. ( B ) Immunofluorescence assays were performed in SMMC-7721/14-3-3ζ cells and CoCl2-treatment in SMMC-7721 cells. The localization of 14-3-3ζ and HIF1α was detected using a Leica confocal microscope. ( C ) CSQT-2/pcDNA3 and CSQT-2/14-3-3ζ were treated with cycloheximide(CHX), and then the expression of HIF-1α and 14-3-3ζ were examined by western blot. ( D ) CSQT-2 and SMMC-7721 cells were co-transfected with HIF-1α and 14-3-3ζ, and then a co-IP assays were performed. Cell-lysates was immunoprecipitated with anti-HIF-1α antibody, and then probed using anti-pan-Ac antibody.

Article Snippet: The pcDNA3 14-3-3ζ plasmid (ID: 9002) was obtained from Addgene (Cambridge, MA, American).

Techniques: Co-Immunoprecipitation Assay, Expressing, Immunoprecipitation, Immunofluorescence, Microscopy, Western Blot, Transfection