sicitk (Santa Cruz Biotechnology)
Structured Review

Sicitk, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/sicitk/product/Santa Cruz Biotechnology
Average 91 stars, based on 2 article reviews
Images
1) Product Images from "Inhibitors of Rho kinases (ROCK) induce multiple mitotic defects and synthetic lethality in BRCA2-deficient cells"
Article Title: Inhibitors of Rho kinases (ROCK) induce multiple mitotic defects and synthetic lethality in BRCA2-deficient cells
Journal: eLife
doi: 10.7554/eLife.80254
Figure Legend Snippet: ( A ) Quantitative real-time PCR of CITK in shBRCA2 HCT116 p21-/- cells transfected with 150 μM of siCITK (N=2). ( B ) Relative cell number (%) of PEO4 and PEO1 after 6 days of being transfected with siROCK (1+2), siCITK or siROCK (1+2)/siCITK and representative images of the transfected cells (N=2). ( C ) Percent of binucleated PEO1 cells transfected with siROCK (1+2), CITK or siROCK (1+2)/siCITK (N=2). ( D ) Quantitative real-time PCR of EMI1 in shBRCA2 HCT116 p21-/- cells transfected with 150 μM of siEMI1 (N=2). ( E ) Relative cell number (%) of PEO4 and PEO1 after 6 days of being transfected with siEMI1 and treated with fasudil (N=2). Representative images of the transfected and treated cells. ( F ) Cell cycle analysis of PEO1 cells following transfection with siEMI1 for 48hs (N=2). Cells were stained with propidium iodide and DNA content was analyzed via FACS (10,000 events per sample). ( G ) Model depicting the events leading to BRCA2-deficient cell death after fasudil treatment. The inhibition or depletion of ROCK in BRCA2-deficient cells leads to cytokinesis failure. As a result, the daughter cells are binucleated (4N) and have extra centrosomes (two instead of one). We speculate that after a subsequent DNA duplication, these cells can attempt mitosis. Mitosis entry with increased DNA content and extra centrosomes may frequently give rise to abnormal and multipolar spindles, leading to misaligned chromosomes and mitotic failure due to multipolar spindle formation. Alternatively, cytokinesis may fail again, and cells may temporarily survive as multinucleated cells, possibly facing cell death during subsequent mitotic attempts.
Techniques Used: Real-time Polymerase Chain Reaction, Transfection, Cell Cycle Assay, Staining, Inhibition
Figure Legend Snippet: ( A ) Relative cell number (%) of shScramble and shBRCA2 HCT116 p21-/- cells at 6 days after transfection with siCITK or siROCK (1+2) (N=2). Representative images are shown on the right. ( B ) Relative cell number (%) of shScramble and shBRCA2 HCT116 p21-/- cells at 6 days after transfection with siEMI1. Samples were treated with fasudil when indicated (N=2). Representative images are shown on the right. Statistical analysis was performed using a two-way ANOVA test followed by a Bonferroni post-test (*p<0.05, **p<0.01, ***p<0.001). Data are shown as the average of independent experiments with the standard error of the mean.
Techniques Used: Transfection