Figure 5.

Axl-148b conjugate promotes necrosis and apoptosis in breast cancer xenotransplants. (A) Scheme of the experiment: Red fluorescent (RFP-expressing) 4175-TGL cells were injected into the mammary gland fat pad of NOD/SCID/IL2R null mice and PBS or axl-148b aptamers were administered into the tumors starting at day 9 post-injection, when masses were palpable, (3 treatments/week, 300 pmol in 100 µl, 10 injections in total, as indicated) and lung metastases were evaluated 11, 18 or 32 days post-tumor-cell injections. (B) FFPE sections of primary tumors were stained with H&E and necrotic areas evaluated: representative images are shown on top of box-and-whisker plots presenting the percentage (%) of necrotic (delimited) versus total areas shown as mean±SEM for the indicated number (n) of mice. (C-D) Primary tumors were stained with Cleaved Caspase-3 (C) or Ki-67 (D) antibodies by IHC and nuclei were counterstained with Hematoxylin (blue): representative images are shown on top of box-and-whisker plots presenting the percentage (%) of positive versus total cells shown as mean±SEM for the indicated number (n) of mice (10 fields/each mouse were evaluated). IHC = immunohistochemistry; H&E = Hematoxilyin & Eosin; FFPE: Formalin-Fixed, Paraffin Embedded; arrows point to positive cells; ns = not significant; * p< 0.05, ** p< 0.01, *** p < 0.001; SEM = Standard Error of Mean; scale bar = 100 µm (B) or 25 µm (C-D).