Stepone software v2.2 rq study

Statistical analysis was done using GraphPad Prism version 9. Data were analyzed by one-way ANOVA following multiple comparison tests to study the statistical significance in different groups. The data obtained from qRT-PCR were evaluated using StepOne Software v2.2 RQ Study (Applied Biosystems/Life Technologies Corp., Carlsbad, CA, USA) and were exported as RQmax and RQmin (2-∆∆Ct +/− ∆∆Ct SD). These results represent the relative quantity of the gene of interest. Conversely, RQmax and RQmin represent the incorporation of the standard deviation of the ∆∆CT into the fold change calculations.

For statistical purposes, cancer Gleason scores were compressed into the 5-tier prognostic grade grouping system endorsed by the International Society for Urologic Pathology and World Health Organizatio12 (link). Gleason 3 + 3 is taken as grade group 1, 3 + 4 is group 2, 4 + 3 is group 3, Gleason score 8 is group 4, and Gleason scores 9–10 are group 5. qRT-PCR data were analyzed using StepOne Software v2.2 RQ Study (Life Technologies Corp.) and exported as RQmax and RQmin (2^{−ΔΔCt +/− ΔΔCt SD}) which represent relative quantities for the gene of interest. RQmax and RQmin are the results of incorporating standard deviation of the ΔΔC_T into the fold-change calculations. Further, statistical analyses on biological replicates of qRT-PCR data were performed with GraphPad Prism 5 software (GraphPad Software Inc.) using one-way analysis of variance (ANOVA) followed by Sidak/Dunnett’s multiple comparison tests. Two-tailed unpaired t-test was done for combined benign and tumor samples.

The LC-MS/MS data from three biological replicates were analyzed using Scaffold_3.6.1 (Proteome Software Inc.). Student's t-test was applied as appropriate way to tell if the abundances are different between the treated (T25) and DMSO control (T0) groups. The results of a t-test are reported as the probability (p-value) that this distance between means could occur by chance. The qRT-PCR data were analyzed using StepOne Software v2.2 RQ Study (Life Technologies Corp.) and exported as RQmax and RQmin (2^{-ΔΔCt +/- ΔΔCt SD}) which represent relative quantity for gene of interest. For the analysis of immunoblots, densitometry was accomplished using Kodak Image Station 4000MM (Carestream Health, Inc., USA) and normalized to β-actin loading control. Further, statistical analyses on biological replicates of qRT-PCR and immunoblot data were performed with GraphPad Prism 5 software (GraphPad Software, Inc., USA) using one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparison test.