Table 1.
RT-PCR primer pairs designed based on sequence information obtained from the Ensembl genomic database (ensembl.org).
Table 2.
Characteristics of patients.
Fig 1.
Schematic representation of the co-culture and wound healing assays with or without macrophages.
(a) Depiction of the co-culture assay. (b) Representative images of the wound healing assay at 0, 8, 16, and 24 h time points. (c) Graphical representation of the wound healing assay results. Data are presented as the mean ± SEM of values from four independent experiments. * p < 0.001, ** p < 0.001.
Fig 2.
Gene expression in ovarian cancer cells and macrophages following co-culture.
(a) mRNA expression of the M2/M1 markers in macrophages following co-culture. (b) mRNA expression of various genes related to chemoresistance and angiogenesis in co-cultured ovarian cancer cells. Immunofluorescence images of (c) single-cultured THP-1 cells, (d) co-cultured THP-1 cells, (e) single-cultured SKOV3-GFP cells, and (f) co-cultured SKOV3-GFP cells. The cyan color represents Hoechst 33342-stained nuclei observed at 440–480 nm with 358 nm excitation. The green color represents the green fluorescence-stained cytoplasm of SKOV3-GFP cells observed at 460–580 nm with 488 nm excitation. Ds Red color represents the fluorescence of PD-L1 observed at 500–700 nm with 480 nm excitation.
Fig 3.
Viability of the single-cultured and co-cultured ovarian cancer cells following carboplatin treatment.
(a) SKOV3 cells co-cultured with THP-1 cells. * p < 0.05, ** p < 0.001, *** p < 0.001. (b) ID8 cells co-cultured with RAW264.7 cells. * p = 0.101, ** p < 0.001, *** p < 0.004.
Fig 4.
Silencing of PD-L1 in human ovarian cancer and macrophage cell lines.
(a) Silencing of PD-L1 in SKOV3 and THP-1 cells. * p = 0.472, ** p < 0.001, *** p < 0.001. (b) Western blotting shows reduced PD-L1 protein expression. (c) Viability of PD-L1-silenced SKOV3 and THP-1 cells after carboplatin treatment. (d) Confocal fluorescence microscopic images of SKOV3 cells.
Fig 5.
Immunohistochemical analysis of PD-L1 expression in ovarian cancer tissues and survival analysis of patients with low and high PD-L1 expression.
(a) Representative immunohistochemical staining of PD-L1. (A) Negative staining, (B) weak staining, (C) moderate staining, and (D) strong staining. Analyses of (b) OS and (c) PFS based on PD-L1 expression.
Fig 6.
Difference in CD68 expression between normal or benign ovarian tissues and ovarian cancer tissues and survival analysis of patients with low and high CD68 expression.
(a) CD68-positive cells in normal, benign, and cancer ovarian cells. * p < 0.05, ** p < 0.01, *** p < 0.001. Analysis of (b) OS and (c) PFS based on CD68 expression.