Figure 4. Inhibition of EYA-PTP activity attenuates tumor growth, vascularization, and cell proliferation.

a) Tumor volume over time from xenografts performed with LLC cells in C57/Bl6 mice and treated with either BZ or vehicle starting when tumor volume was >100 mm³ (arrow). 25 μg/g body weight BZ or vehicle was delivered intra-peritoneally. Tumor growth was monitored by caliper measurements until the experiment was terminated. BZ treatment substantially reduced tumor size. Data represented as mean and SD; two-way ANOVA with a Bonferroni’s post-test was used. (n = 9 (vehicle), n = 8 (BZ), *** P<0.001).

b) Final tumor weights from xenografts performed with LLC cells in C57/Bl6 mice and treated with either BZ or vehicle. Data represented as a box and whiskers plot where the box extends from the 25^th to 75^th percentile and the whiskers are drawn between the smallest and largest values. The median is indicated by a horizontal line in the box. (n = 9 (vehicle), n = 8 (BZ), unpaired t-test).

c) Tumor volume over time from xenografts performed with LLC cells in C57/Bl6 mice and treated with either BZ or vehicle starting when tumor volume was >600 mm³ (arrow). 25 μg/g body weight or vehicle was delivered intra-peritoneally. Tumor growth was monitored by caliper measurements until the experiment was terminated. BZ treatment substantially reduced tumor size. Data represent mean and SD (n = 7 (vehicle), n = 6 (BZ), ** P<0.01, *** P<0.001).

d) Vehicle and BZ treated tumors co-stained with the endothelial marker CD31 and the proliferation marker Ki-67. Sections were counter-stained with DAPI to show nuclei. Scale 100 μm.

e) Quantitation of CD31-positive area in Vehicle and BZ treated tumors. Four representative images were analyzed from each tumor. Data represents mean and SD (n = 5 tumors in each group).

f) Quantitation of Ki-67 positive cells in LLC and EYA3-ko tumors. Four representative images were analyzed from each tumor. Data represents mean and SD (n = 4 tumors in each group).

g) Quantitation of TdT-mediated dUTP nick-end labeling (TUNEL) staining in LLC and EYA3-ko tumors. Four representative images were analyzed from each tumor. Data represents mean and SD (n = 3 tumors in each group).

h) Tumor volume over time from xenografts performed with LLC Eya3-ko cells in C57/Bl6 mice and treated with either BZ or vehicle starting when tumor volume was >600 mm³ (arrow). 25 μg/g body weight BZ or vehicle was delivered intra-peritoneally. BZ treatment had no effect on tumor size. Data represented as mean and SD (n = 7 (vehicle), n = 6 (BZ)).