Figure 2

Whole mount immune fluorescence staining for Ki67. Vybrant CM-DiI labeled LGSOC cells (red), nuclei stained with DAPI (blue) and anti Ki-67 (green). (a–d) The number of xenografts analyzed for Ki67 is indicated in the figures; all images are at 25X magnification (scale bar 50 µM). Tumor area is indicated with a white line. (e–h) Quantification of Ki67 expression in zebrafish xenografts using ImageJ. Generation of Maximum Intensity Projections (MIPs) from relevant Z-stacks (e), transformation of 8-bit files to RGB file (f), binary masking of Ki67 staining using fixed threshold (g), select ROI of tumor (h), percentage of stained area in ROI represents Ki67 expression. (i) Percentages of different treatment conditions are presented as AVG ± SEM and each dot represents a xenograft. Multiple comparisons between treatment groups were determined by One-way ANOVA; Tukey procedure was used to adjust p-values. p-values of 0.05 were considered as statistically significantly different and output was represented as: not-significant (ns) p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, and **** p ≤ 0.0001.