FIGURE 7.

DAXX and DAPK3 are inversely correlated in vivo. A, quantitative immunohistochemical analyses were performed using excised tumor tissue corresponding to CNTL, DAXX K/D, and DAPK3 K/D tumors from ALVA-31 subcutaneous injections, as described above. Tissue sections were stained using an antibody specific for DAPK3. A negative control (hematoxylin) was also included. Representative ×10 images are shown. Immunostaining was quantified and analyzed using Aperio ImageScope software, and the graph was constructed using GraphPad Prism version 5 software. An inverse correlation between DAXX and DAPK3 is observed. B, immunostaining using LC3 or p62 antibodies shows increased autophagy in DAXX K/D tumors, based on LC3 increase and p62 decrease in these tumors. The pattern is reversed for DAPK3 K/D tumors. Representative ×40 images are shown. C, immunohistochemistry using CD31 marker for angiogenesis shows statistically significant increased proliferation in larger size tumors (DAPK3 K/D) compared with smaller ones (DAXX K/D). Quantification was done as in A above. Error bars, S.E.