Kiss1r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Functional genomics identifies novel genes essential for clear cell renal cell carcinoma tumor cell proliferation and migration"
Article Title: Functional genomics identifies novel genes essential for clear cell renal cell carcinoma tumor cell proliferation and migration
Figure Legend Snippet: Decreased KISS1R, KSR1, CAMK1, and SSPN expression results in loss of invasive phenotype in ccRCC cells (A) Images represent 10x magnification of NT and specified hit knockdown cells plated in a 3D matrix after 10 days of growth. (B) 40x manual magnification of representative colonies of NT and target hit knockdown cells grown in 3D cell culture assays at day 10. Stellar outgrowth is seen in NT populations and is indicated by black arrows. (C) Invasion assays of NT and target hit knockdown cells. Representative 10x images of transwell inserts from invasion chambers are shown. Invasion is quantitated as number of invading cells per visual field (n=3). (D) Immunofluorescence for phalloidin, VASP, and dapi (nuclear stain) in NT and hit target knockdown cells. Top panels for each cell line are 20x magnification. Highlighted areas magnified manually, and are shown directly below panel of origin.
Techniques Used: Expressing, Cell Culture, Immunofluorescence, Staining
Figure Legend Snippet: Functional Validation of tumor specific expression of a subset of hits identified in RCC (A) Box whisker plot of QPCR evaluating CAMK1, KISS1R, KSR1, SSPN, and CDH13 mRNA expression in stage I patient tumor and matched normal samples (n=8 for each normal and tumor samples). Results are shown as tumor fold change induction vs. normal samples, where the dashed line represents normal tissue expression. (B) Western blot for CAMK1 and SSPN in protein lysates derived from stage I-IV patient tumor and matched normal samples. Protein levels are quantitated against β-actin loading control, and tumor samples are normalized against matched normal samples which are set at 1. Samples are further sorted by stage, and tumors that demonstrate over a 20% increase in expression vs. matched normal are considered to be significantly overexpressed(*). (C) IHC of patient tissue microarrays for protein expression of KISS1R (normal n=40, 24, 25, 6, 9 and tumor n=36, 15, 23, 6, 20 for stages I, II, III, IV, and metastasis, respectively), KSR1 (normal n=53, 34, 34, 8, 8 and tumor n=42, 21, 23, 6, 24 for stages I, II, III, IV, and metastasis, respectively), and CDH13 (normal n=53, 29, 33, 8, 9 and tumor n=40, 19, 22, 8, 19 for stages I, II, III, IV, and metastasis, respectively). Representative tumor and normal images as well as mean H-scores ± standard deviation are shown for KISS1R and KSR1; mean I-scores ± standard deviation are shown for CDH13. Asterisk indicates values of statistically significant increases in tumor samples (where p≤0.05).
Techniques Used: Functional Assay, Expressing, Whisker Assay, Real-time Polymerase Chain Reaction, Western Blot, Derivative Assay, Immunohistochemistry, Standard Deviation
Figure Legend Snippet: Decreased expression of KISS1R, KSR1, CAMK1, and SSPN leads to tumor cell death and/or senescence (A) Protein expression analysis of KISS1R, KSR1, CAMK1, and SSPN in NRE and ccRCC cell lines via western blot. VHL mutation status is provided. (B) QPCR for mRNA transcript expression in NT and hit knockdown cells are presented as fold change relative to NT control, where NT values are set at 1. (C) Proliferation of NT vs. hit targeted shRNA cell lines. (D) Western blot for protein expression in NT control and shRNA targeted cells for PARP, and P21. β-actin was used as a loading control. (E) Flow cytometry analysis of propidium iodide stained NT and target knockdown cells evaluating cell death. Increases of 5% or greater versus NT control cells were considered to be significant.
Techniques Used: Expressing, Western Blot, Mutagenesis, Real-time Polymerase Chain Reaction, shRNA, Flow Cytometry, Cytometry, Staining