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. 2021 Sep 16;12:5356. doi: 10.1038/s41467-021-25553-z

Fig. 4. High PRC1 expression levels prime EwS cells for PLK1 inhibition.

Fig. 4

a Heatmaps depicting the average percentage of growth inhibition in EwS cells with/without PRC1 silencing after 72 h of PLK1 inhibition; n = 6 biologically independent experiments. b Analysis of IC50 in CRISPR Cas9-edited A673 cells after 72 h of PLK1 inhibition. Data are displayed as individual dots. Horizontal bars represent means and whiskers SEM, n = 6 biologically independent experiments. c Sphere-formation of EwS cells treated with BI2536 or BI6727. Data are displayed as individual dots. Horizontal bars represent means and whiskers SEM, n = 9 biologically independent experiments. Representative images are shown; scale bar = 50 µm. d Analysis of apoptosis of EwS cells with/without PLK1 inhibition for 72 h. Data are displayed as individual dots. Horizontal bars represent means and whiskers SEM, n = 6 biologically independent experiments. e Schematic of PLK1 inhibitor treatment in vivo. f Individual tumor volume curves (n = 6 per condition for NC-cells, n = 7 per condition for KO-cells). Representative micrographs of H&E and PRC1, the positivity of cleaved caspase-3 (CC3), as well as γ-H2AX per HPF. Scale bar = 100 µm. g Excess over Bliss analysis of RDES cells with/without PRC1 knockdown treated with combinations of Vincristine (VCR) and the PLK1 inhibitors in vitro (red indicates synergy). Summary level (mean) data from n = 6 biologically independent experiments. h Analysis of tumor volume of mice treated with either vehicle, VCR (1 mg/kg/d, i.p.) alone, BI6727 (5 mg/kg, i.v.) alone, or in combination. Representative micrographs of PRC1 staining and positivity of γ-H2AX, CC3, and Ki-67 per HPF. n = 6 biologically independent animals per condition. Scale bar = 100 µm. i Dose-response curves of two Doxorubicin (Doxo)-resistant EwS cell lines (A673, TC71) treated with Doxo combined with BI2536 or BI6727 for 72 h. The reverse index (RI) is given. Dots represent means and whiskers SEM, n = 6 biologically independent experiments. j Median effective dose (ED50) for Doxo-treatment alone or in combination with PLK1 inhibitor BI2536 or BI6727 for 72 h in Doxo-resistant A673 and TC71 cells. Data are displayed as individual dots. Horizontal bars represent means and whiskers SEM, n = 6 biologically independent experiments. k Schematic illustrating key findings of this study. Two-sided Mann-Whitney test in all panels.