Journal: eLife
Article Title: A single cysteine residue in vimentin regulates long non-coding RNA XIST to suppress epithelial–mesenchymal transition and stemness in breast cancer
doi: 10.7554/eLife.104191
Figure Lengend Snippet: ( A ) Morphology of MCF-7-expressing WT-VIM and C328S-VIM in brightfield (scale bar = 100 µm). ( B ) Morphology of WT and C328S cells stained with CellMask deep red dye. Images were captured by INCA 2200 and analysed using INCarta software (scale bar = 50 µm). ( C ) Differences in nuclear area, nuclei form factor, nuclear major axis, nuclear minor axis, cell area, cell diameter, cell perimeter, cell minor axis, cell major axis, and cell form factor between the two cell lines. ( D ) Significant reduction in nuclear compactness and nuclei/cell area between the two cell lines. Proliferation rate was compared between WT and C328S cells by ( E, F ) colony, ( G ) MTT, and ( H ) CyQUANT assays. ( I ) CyQUANT cell adhesion assay was performed to compare the cell adhesion between WT and C328S cells without substrate, and ( J ) with the addition of laminin, fibronectin, and collagen, separately. ( K ) Chemotactic migration of the WT and C328S cells through 8.0 µm culture inserts. The cells were fixed and stained with 0.1% (w/v) crystal violet before imaging. ( L ) The cells on the outer surface of the inserts were counted and compared between WT and C328S. ( M ) Chemotactic invasion in WT and C328S cells through 8.0 µm culture inserts coated with Matrigel. The cells were fixed and stained with 0.1% (w/v) crystal violet. ( N ) Total number of cells invaded on the outer surface of the inserts was counted. Statistical analyses: n = 3, error bars = ± SEM, Student’s t -test was used to calculate p values using Microsoft Excel and are given by asterisks (*p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001).
Article Snippet: Colour development was achieved using 3, 3’-diaminobenzidine (DAB) substrate, and slides were lightly counterstained with Harris haematoxylin, dehydrated, mounted in Permount, and imaged by brightfield microscopy (Keyence Corporation, Itasca, IL).
Techniques: Expressing, Staining, Software, CyQUANT Assay, Cell Adhesion Assay, Migration, Imaging