Computational protocol: p21-activated kinase 1 determines stem-like phenotype and sunitinib resistance via NF-κB/IL-6 activation in renal cell carcinoma

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Protocol publication

[…] ALDH activity was detected using the ALDEFLUOR assay kit (Stem Cell Technologies, Vancouver, BC, Canada) as described by the manufacturer. Briefly, dissociated single cells from cell lines were suspended in ALDEFLUOR assay buffer containing an ALDH substrate, bodipy-aminoacetaldehyde (BAAA), at 1.5 μM and incubated for 1 h at 37°C. A specific inhibitor of ALDH, diethylaminobenzaldehyde (DEAB), at a 10-fold molar excess, was used as negative control. Flow cytometry data were analyzed by FlowJo software (TreeStar, Ashland, OR, USA). [...] Experimental data were presented as mean±S.D. Student's t-test was used to compare continuous variables. The χ2 test or Fisher's exact test was used to compare qualitative variables. Survival was calculated from the date of surgery to the date of death or last follow-up. Kaplan–Meier method with log-rank test was applied to compare survival curves. All statistical tests were two sided and performed at a significance level of 0.05. Univariate and multivariate Cox regression models were used to analyze the impact of prognostic factors. The relationships between variables were analyzed using Spearman's correlation coefficient for correlation and χ2 test for trend. Data were analyzed using SPSS Statistics 21.0 (SPSS Inc., an IBM Company, Chicago, IL, USA) and GraphPad Prism5 (GraphPad Software, La Jolla, CA, USA). […]

Pipeline specifications

Software tools FlowJo, SPSS
Applications Miscellaneous, Flow cytometry
Organisms Homo sapiens
Diseases Carcinoma, Renal Cell, Neoplasms, Cardiovascular Abnormalities