Validation of KMT2D function in MM23, MM2, and MM25 PDXs

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Validation of KMT2D function in MM23, MM2, and MM25 PDXs Validation of KMT2D function in MM23, MM2, and MM25 PDXs. A–D, MM23 (NRAS mutated) or MM2 and MM25 (BRAF mutated) PDX2 cells expressing control (Luc) or KMT2D (pool of two) shRNAs were injected in recipient mice (B), or plated for a luminescent-based prolif... Validation of KMT2D function in MM23, MM2, and MM25 PDXs. A–D, MM23 (NRAS mutated) or MM2 and MM25 (BRAF mutated) PDX2 cells expressing control (Luc) or KMT2D (pool of two) shRNAs were injected in recipient mice (B), or plated for a luminescent-based proliferation (C) or Transwell migration (D) assays. A, qPCR analysis of KMT2D mRNA levels in PDX2-silenced cells at time of transplantation or plating of in vitro assays. The RPLP0 mRNA level was used as housekeeper. B, in vivo melanoma growth in NSG mice. Tumor volumes (mean ± SD) of PDX melanomas were measured after 6, 5, and 4 weeks in MM23, MM3, and MM25, respectively. C, relative migration (mean ± SD) of PDX-silenced cells calculated as described in Fig. 2. D, relative growth of PDX2-silenced cells after 3 days of culture. Values (mean ± SD) are expressed as a ratio of the mean proliferation value in the silenced cells compared to control (Luc shRNA) cells. P value is calculated by the Student t test (*, P < 0.01). Daniela Bossi et al. Cancer Discov 2016;6:650-663 ©2016 by American Association for Cancer Research