The relative quantification value, fold difference, is expressed as 2-ΔΔCt. Statistical analysis Statistical analysis was performed with MedCalc Software, Version 11.3.2 (Mariakerke, Belgium). All values were expressed Blasticidin S purchase as Median ± Interquartile Range (IQR) because a normal distribution of gene and protein expression could not be confirmed by the D’Agostino-Pearson test. Therefore, the Median value was chosen to divide patients in two different groups. Survival time was determined as the time from tumor resection to tumor conditional death and as the time from tumor resection to time of obvious recurrence. The overall
survival (OS) time in association with LgR5 expression was estimated using the Kaplan-Meier method . To analyze
differences in the overall/tumor related survival among patients after successful (R0) curative surgical resection for EAC patients were divided into two subgroups (dichotomous variables). Median cut-off value for either high or low expressors was set at 33% for LgR5 expression in BE (n = 41), 15% for LgR5 expression in adjacent EAC (n = 41), and 15% for LgR5 expression in all EAC (n = 60); univariate Tozasertib mw analysis of significance for LgR5 expression differences in survival curves was evaluated with the log rank test. Multivariate with the Cox Proportional Hazards Model  was performed including all parameters that were found to be significant on univariate analysis. Fisher’s exact test was used to investigate the Palbociclib price relation between two categorical variables. Data were analyzed using the non-parametric Mann-Whitney U test or Kruskal-Wallis test when more than 2 groups were compared. P values of less Aldehyde dehydrogenase than 0.05 were regarded statistically significant. Results LgR5 Immunohistochemistry Immunohistochemistry against the putative intestinal stem cell marker LgR5 showed
positive stainging in 85% (35 of 41) of the specimen of patients with EAC with BE, and 84% (16 of 19) in EAC without BE (p = n.s). No LgR5 expression was found in specimen with esophageal SCC. No expression of the putative stem cell marker (LgR5) was detected in normal esophageal squamous cell epithelium, adjacent to the tumor. Normal colon mucosa (used as positive control) showed the typical staining pattern of LgR5 (Figure 1a and 1b), as they stained the well-described putative colon mucosa stem cells, located at the basis of the crypts, or the transit amplifying zone, which are regarded to resemble the stem cell niche [24, 25]. Figure 1 Immunohistochemical staining of LgR5 (membranous staining pattern, brown) in normal colon tissue. Normal colon mucosa (asterisk) showed the typical staining pattern as they stained the well-described putative colon mucosa stem cells, located at the basis of the crypts, or the transit amplifying zone, which are regarded to resemble the stem cell niche (arrows).