## 3 Results and Discussion
### Analysis of Apoptosis Using the Muse(r) Flow Cytometer  
Flow cytometry with the Muse(r) Annexin V & Dead Cell Assay, Muse(r) MultiCaspase Assay Kit, and Muse(r) Bcl-2 Activation Dual Detection Assay Kit was used to investigate the mode of studied juices-induced cell death in DU145 and LNCaPP prostate cell lines. The Muse(r) Annexin V & Dead Cell Assay for quantitative analysis of live, early/late apoptotic, and dead cells was used. Annexin V is a calcium-dependent cytoplasmic protein with a high affinity for phosphatidylinositolserine, inner cell membrane component. During apoptosis, annexin V can bind phosphatidylserine, which translocates to the outer surface of the cell membrane [26]. In this research, we also performed multi-caspase activity (caspase-1, -3, -4, -5, -6, -7, -8, and -9) to confirm the apoptosis mechanism. The Muse(r) MultiCaspase Assay Kit determines the count and percentage of cells with caspase activity, in combination with a dead cell dye. The detection assays revealed a significant (\(p\leq 0.05\)) decrease in live cells in the examined juices treatment in comparison to the medium control in DU145 and LNCaPP prostate cancer cell lines (Tables 2-5; Supplementary Figures S1 and S2). Additionally, there was a significant (\(p\leq 0.05\)) increase in the total apoptosis rate and number of caspase positive cells compared to the untreated control in both cell lines (Tables 2-5; Supplementary Figures S1 and S2). In DU145 and LNCaPP cell lines, after treatment of the young shoots' juice that was subjected digestion and absorption, the total apoptosis rate was 44.50 \(\pm\) 2.83% and 55.37 \(\pm\) 0.57, respectively (Tables 2 and 4; Supplementary Figures S1 and S2). After treatment with juices, the rate of early apoptosis exceeded that of the late, however, for the young shoots' juice subjected to in vitro digestion and absorption in the LNCaPP cell line, late apoptosis was predominant (Tables 2 and 4; Supplementary Figures S1 and S2). We found that the number of caspase positive cells was increased in DU145 and LNCaPP cell lines and this activation depended on the juice used (Tables 3 and 5;  
Figure 1: Effect of juice of young shoots of red cabbage (ys d + a) and of the mature vegetable (mv d + a) subjected to in vitro gastrointestinal digestion and absorption on DU145 and LNCaPP prostate cancer cells and PNT-2 normal cancer cells proliferation in test with using Cell Proliferation ELISA, BrdU (Sigma-Aldrich). Values are expressed as means \(\pm\) SD for \(n\) = 9, standardized to an untreated control set as 100%. Statistical significance was based on \(t\)-test * \(p\leq 0.05\). Supplementary Figures S1 and S2). It was observed that young shoots' juice subjected to in vitro gastrointestinal digestion and absorption increased the number of caspase positive cells more effectively in comparison to the digested and absorbed juice of the mature vegetable in both cell lines (Tables 3 and 5; Supplementary Figures S1 and S2). Similar results were noticed for the juice of young shoots not subjected to the aforementioned processes (Tables 3 and 5; Supplementary Figures S1 and S2). Similar to our study, it has been stated that bioaccessible fractions of broccoli, mustard, and radish microgreens show induction of early apoptosis in Caco-2 cells [27]. Similarly, extracts of fresh green cabbage, compared with the control group, induced cell apoptosis in A529 cells by increasing early apoptosis [28]. [PERSON] et al. [29] showed that red cabbage extract inhibits HeLa and HepG2 cancer cell lines growth through the induction of apoptosis via caspase-dependent pathway. According to [PERSON] et al. [30], glucosinolates induce caspase-mediated apoptosis in cultured PC-3 human prostate cancer cells. \begin{table}