Fig. 7
The ESCRT-II complex in luminal epithelial cells is under the influence of embryonic signals. A Schematic representation of the ESCRT complex and Ras-related proteins involved in EV biogenesis and trafficking. B Diagram showing possible positions of binding motifs for estrogen (ERE, blue box) and prostaglandin (PGRE1, red; PGRE2, green; PGRE3, purple boxes) response elements upstream/downstream from the VPS25 and VPS36 TSS. C–F mRNA expression of VPS22, VPS25, VPS36, and RAB27A in response to treatment with E2 (100 nM) or PGE2 (100 nM) in primary luminal epithelial cells. Cells were pretreated with specific inhibitors – MPP for ESR1 (1 h, 1 µM) or AH6809 for PGE2 (10 min, 10 µM). Gene expression was normalized to HPRT1 (AU), identified as the best reference gene by the NormFinder algorithm. Data for separate E2 and PGE2 treatments were analyzed using one-way ANOVA with Tukey’s multiple comparison test (VPS25, VPS36, RAB27A; D, E, F; p < 0.05) or Kruskal–Wallis and Dunn’s multiple comparison tests (VPS22; C), and are shown as means ± SEM (vs. control; n = 4–7). Data presenting simultaneous treatment with both embryonic signals were analyzed using paired T-test (C, D, and F; p < 0.05) or Wilcoxon matched-pairs signed rank test (E; p < 0.05) and are presented as individual plots (control-treatment). AU – arbitrary units, TSS – transcription start site