Computational protocol: mTOR inhibition improves fibroblast growth factor receptor targeting in hepatocellular carcinoma

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Protocol publication

[…] Western blotting was performed as described previously (). Briefly, whole-cell lysates were prepared and protein samples (50 μg) were subjected to western blotting on a denaturating 10% SDS–PAGE. Membranes were probed with indicated antibodies and detected by chemiluminescence (Amersham Bioscience, Piscataway, NJ, USA). To quantify results from western blotting, densitometry was performed using ImageJ (1.46r; http://imagej.nih.gov.ij). Expression of proteins was calculated in relation to the respective control (e.g. pAkt/Akt). [...] Statistics were performed using SigmaStat (Version 3.0, San Jose, CA, USA) and GraphPadPrism (Version 6.0, La Jolla, CA, USA). Results of in vivo experiments were analysed for significant outliers using Grubb's test (http://www.graphpad.com). Tumour-associated variables in in vivo experiments were tested for significance using the Mann–Whitney U-test and ANOVA test. The two-sided Student's t-test was applied for analysis of in vitro data. Results for migration assays and PCR are shown relative to control results. Expression of FGFRs in all HCC cell lines, stromal cells and patient samples is normalised to Hep3B as this tumour cell line expresses all FGFRs. All results were confirmed in independent experiments and are expressed as the mean±s.e.m. […]

Pipeline specifications

Software tools ImageJ, GraphPad Prism
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Mus musculus, Homo sapiens
Diseases Carcinoma, Hepatocellular, Neoplasms
Chemicals Sirolimus