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. 2015 Aug 24;8(4):295–307. doi: 10.1016/j.tranon.2015.06.002

Figure 5.

Figure 5

CDK5 knockdown prevents metastatic melanoma spread in murine dissemination models. (A) Formation of lung and liver metastases after intravenous injection of SKMel28 melanoma cells in immunodeficient HPRT NOD-SCID mice was determined. In this model system, doxycycline-induced shRNA-mediated CDK5 knockdown led to a dramatic reduction in the average number and size of pulmonary lesions as compared with uninduced controls (left panel). Liver metastases were completely abolished upon induced CDK5 knockdown, whereas they were readily observed in uninduced controls (right panel). The figures show representative H&E stains of FFPE tissue sections. (B) Injection of B16 murine melanoma cells into the tail veins of C57BL/6 wild-type mice was applied as a third, syngenic in vivo model system to validate the effect of CDK5 abrogation on melanoma metastasis. One of five CDK5-specific shRNA-clones (“CDK5#1”) showed reproducible knockdown of CDK5 in B16 murine melanoma cells as compared with scrambled controls using Western blot analysis. (C) When this clone CDK5#1 was used in a tail vein injection model, a dramatic decrease in both the amount and average size of lung colonies representing pulmonary metastases was observed upon CDK5 knockdown as compared to mock-transfected cells (macroscopic image from two representative animals).