FIGURE 10:

Rac1 overexpression increases tumor burden. OVCAR8 GFP control, OVCAR8 GFP-Rac1 OE, and SKOV3ip CRISPR-Cas9 control, SKOV3ip Rac1 CRISPR-Cas9 KD clone ovarian cancer cells transduced with luciferase were IP injected into NSG mice. Tumor burden was measured using bioluminescent IVIS imaging. (A) Representative IVIS images of NSG mice injected with OVCAR8 GFP control (top row) and OVCAR8 GFP-Rac1 OE cells (bottom row) imaged 2 and 4 wk post–IP injection. (B) Quantification of the average radiance at 2 and 4 wk postinjection. Quantification of 2D radiance using two-way ANOVA shows significant time-dependent variance; p = 0.0043 of tumor burden. N = 4 for each group. (C) Representative IVIS images of NSG mice injected with SKOV3ip CRISPR-Cas9 control cells or Rac1 CRISPR-Cas9 KD cells imaged 18 h and 1 wk post–IP injection. (D) Quantification of the average radiance at 18 h and 1 wk post-injection. Quantification of average radiance using two-way ANOVA reveals a significant decrease in tumor burden in mice injected with CRISPR KD clones (clones 26 and 27) compared with control. N = 5 for each group. *p < 0.05, **p < 0.01. (E) Omental tissue isolated from mouse xenografts of SKOV3ip ovarian cancer cells expressing luciferase. Omental tissues were isolated from mice 1 wk after being injected with 4 × 10⁶ Rac1 CRISPR-Cas9 KD (C3-26 or C3-27) or CRISPR-Cas9 control (C4) cells, and gene expression levels of luciferase were analyzed based on targeted qPCR. Data represented are mean ± SE. These data are combined from one experiment with five biological replicates. **** indicates p value ≤ 0.0001, where values represent relative expression for Rac1 CRISPR-Cas9 KD compared with CRISPR-Cas9 control C4 (1.0) and normalized to 18s rRNA using unpaired two-tailed t test.