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. Author manuscript; available in PMC: 2023 Feb 3.
Published in final edited form as: Mol Cancer Res. 2023 Feb 1;21(2):170–186. doi: 10.1158/1541-7786.MCR-22-0486

Figure 4. TWEAK in combination with carboplatin enriches for CSCs and eliminates non-CSCs.

Figure 4.

(A) Stem cell marker flow cytometry of HGSOC cells treated with vehicle, TWEAK (100ng/ml), carboplatin (125μM), or TWEAK and carboplatin for 72 hours. Representative gating for CAOV4 (left), percent positive cells representing mean plus SD (right). One-way ANOVA, Tukey’s post-hoc test (n=3). (B) qRT-PCR for select stemness genes in HGSOC cells with vehicle, TWEAK (100ng/ml), carboplatin (125μM), or TWEAK and carboplatin for 72 hours represented relative to vehicle. One-way ANOVA, Tukey’s post-hoc test (n=3). (C) Spheroid efficiency of HGSOC cells treated with vehicle, TWEAK (100ng/ml), carboplatin (125μM), or TWEAK and carboplatin for 4 days represented relative to vehicle. One-way ANOVA, Tukey’s post-hoc test (n=4). (D) Flow cytometry analysis of CD117 expression on Fn14+ cells treated with vehicle, TWEAK (100ng/ml), carboplatin (125μM), or TWEAK and carboplatin for 72 hours. Representative gating (left) and percent positive cells representing mean plus SD (right). One-way ANOVA, Tukey’s post-hoc test (n=3). (E) Viability of sorted OV90 CD117+ or CD117− cells grown for 8 days normal media (top). Two-way repeated measures ANOVA, Sidak’s post-hoc test (n=4). Spheroid efficiency of OV90 CD117+ or CD117− cells treated with carboplatin (90μM) for 4 days represented relative to vehicle (bottom). Unpaired t-test (n=4). (F) qRT-PCR of select genes for sorted CD117+ or CD117− cells treated with vehicle or TWEAK (100ng/ml) for 72 hours. One-way ANOVA, Tukey’s post-hoc test (n=4). (G) Viability of CD117+ sorted CAOV4 cells treated with either Carboplatin (125μM), Carboplatin and TWEAK (100ng/ml), TWEAK then Carboplatin, and Carboplatin then TWEAK for 120 hours. One-way ANOVA, Tukey’s post-hoc test (n=3). (H) Flow cytometry analysis of CD117 and Fn14 expression CAOV4 cells mock sorted or CD117− sorted cells treated with vehicle or TWEAK (100ng/ml) and carboplatin (30μM) for two cycles of 7 days, where initial treatment is for 24 hours with subsequent two-fold dilution with media. Representative gating (top) and percent Fn14+CD117+ cells representing mean plus SD. Two-way ANOVA, Sidak’s post-hoc test (n=4 mock, 5 CD117− sorted). Data represent mean and SEM. * p<0.05.