Figure 2.

Targeting CD47 decreases melanoma tumor burden in vivo. (A) Schematic of the in vivo syngeneic mouse melanoma treatment regimen. C57Bl/6 mice were injected subcutaneously with B16 melanoma cells into the hind flank. Once tumors reached 100 mm³, intraperitoneal injections of 10 uM CD47 morpholino (CD47(-)) or saline began once a week for 3 weeks. (B) Tumor volume was determined throughout the study (LW2/2). (C) Mice were euthanized at the end of the study (day 21) or when tumor volume reached 1500 mm³ where tumor weight was determined (*p<0.05, n=6–8/group). (D) Representative PAI imaging for tumor sO₂ along with Power Doppler in 3D-Mode to determine tumor vascularity. (E) Tumor sO₂ was quantified with the Vevo 2100 LAZR software tools while (F) tumor vascularity was quantified with Vevo CQ software of WT and CD47 targeted tumors (*p<0.05, n=4–5/group). (G) Representative immunohistochemistry images of tumor sections for (H) carbonic anhydrase, (I) intratumoral cytotoxic (yellow cells due to colocalization of CD3+ (red, APC) and CD8+ (green, FITC) T cells and (J) granzyme B (*p<0.05, n=4–5/group). Images were obtained at 20x magnification with the Olympus BX43 microscope and quantified using the PerkinElmer Mantra and inform analysis. (K) Gene expression of perforin-1 was determined through RT-qPCR of tumors (*p<0.05, n=4–5/group). PAI, photoacoustic imaging; WT, wild type.