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. Author manuscript; available in PMC: 2020 May 13.
Published in final edited form as: Oncogene. 2019 Nov 13;39(8):1619–1633. doi: 10.1038/s41388-019-1097-7

Figure 1: Ovarian cancer SIRT3 expression is context-dependent, and increases in response to anchorage-independence.

Figure 1:

A. SIRT3 mRNA expression in tumor samples from primary tumors of the ovary, matching ascites, and omental and peritoneal metastatic lesions (Geo:GSE85296, n=4, mean expression at each site per patient shown, repeated measures ANOVA P = 0.047, Dunnett’s multiple comparison test **P = 0.005). B. Ovarian cancer cell lines were cultured in anchorage-independent (a-i) conditions using ultra low attachment (ULA) plates for 72 h, and SIRT3 protein expression compared to cell cultures in attached (A) conditions using western blotting. Densitometry of SIRT3 bands is expressed relative to loading control GAPDH, and levels in a-i compared to attached conditions for each cell line (n=3–6; t-test ***P<0.001, ****P<0.0001). C. Epithelial ovarian cancer cells (EOCs) were derived from ascites of Stage III and IV high grade serous adenocarcinoma patients and cultured in attached or a-i conditions for 72 h. SIRT3 expression was assessed as in B (n=5, paired t-test **P=0.07). D. mRNA levels of sirtuin family members were assessed using U133 microarray after EOCs derived from patient ascites (n=5) were cultured in a-i conditions for 72 h as described in [47]. mRNA levels are expressed relative to cells grown in attached conditions. E. SIRT3 protein expression was assessed by western blotting of lysates from attached cell cultures (A), cells maintained for 24 h in ULA plates (a-i) and 24 hours following re-attachment (re-A). Densitometry of SIRT3 bands is expressed relative to loading control actin and normalized to attached conditions (n=3; one-way ANOVA, Tukey’s multiple comparison test *P<0.05, **P<0.01). F. SIRT3 mRNA expression was assessed by semi-quantitative real time RT-PCR following cell culturing as in E, and expressed relative to OVCA420 in attached conditions (n=3; two-way ANOVA, Tukey’s multiple comparison test *P<0.05, **P<0.001, ****P<0.0001).