Fig. 5

ER stress in HGSOC and adjacent ovarian tissues. (A) Heatmap summarizing the outcomes of immunohistochemical staining of a subset of paraformaldehyde-fixed HGSOC tumors. The data are shown for the quantity of positive cells among cells of the indicated type. (B, C, D) Examples of immunohistochemical staining for BiP (200× magnification) in tumor tissue (B), and tissue from the adjacent ovary (C); a negative control of the tumor tissue is also provided (D). (E) Changes in the relative cell count of tumor cells (tumor) following incubation with TUDCA in complete medium (+Q, +G) or medium with depleted glucose (+Q, -G). (F) Changes in BiP levels following treatment with compounds that modulate ER stress. The tumor cells (tumor) and ovarian fibroblasts (fibroblasts) were incubated with DMSO (as a control), salubrinal, copper(II)-phenanthroline complexes C0 and C1, a mixture of C1 with salubrinal, or tunicamycin, and the protein expression of BiP was subsequently analyzed by Western blotting. The heatmap shows the relative expression levels of BiP in individual patients. The data are shown as log2 quantities relative to the DMSO control (0; white). Blue indicates decreased expression; red indicates increased expression. (G-H) Changes in the resazurin reduction rate and cellular ATP levels following treatment with compounds that modulate ER stress. The data are shown for the alamarBlue and CellTiter-Glo assay measurements, which were analyzed as values relative to those of tumor cells cultivated in a complete medium. The cells were incubated with DMSO (as a control), salubrinal, copper(II)-phenanthroline complexes C0 and C1, a mixture of C1 with salubrinal, or tunicamycin. The data from three independent tumor cell isolates, each measured four times, are shown