Dissemination of NHL is dependent on CD47. (A) Raji cells were transduced with lentiviruses encoding shRNA CD47 knockdown constructs (shCD47) or a GFP control, and CD47 protein expression was determined by flow cytometry. (B) Relative CD47 expression levels were quantified by comparing mean fluorescence intensity with wild-type Raji cells. (C) Raji cells from panels A and B were transplanted subcutaneously into the right flank of adult NSG mice and monitored weekly for tumor growth. Representative mice are shown 28 days after transplantation with flank tumors demarcated (red box). (D) Tumor volume was quantified for all mice (n = 5 per cohort), demonstrating reduced growth from shCD47-1,2–transduced cells compared with GFP control. Statistical comparison was performed using a 2-way ANOVA. Data are mean ± SD. (E) Posttransplantation day 28, these mice were killed and analyzed for the number of gross liver metastases with representative mice shown (livers are shown in right panels). Direct invasion from the primary tumor (black arrows) to adjacent organs was not observed. (F) Number of liver metastases was quantified for all mice, with each data point representing an individual mouse. P values were calculated by Student t test: **P < .005. ***P < .0005.