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. Author manuscript; available in PMC: 2017 May 14.
Published in final edited form as: Oncogene. 2016 Nov 14;36(8):1080–1089. doi: 10.1038/onc.2016.272

Figure 2. Synthetic lethality of Y-27632 with VHL loss is mimicked by siRNA downregulation of ROCK1, not ROCK2.

Figure 2

RCC4±VHL matched cell lines were transfected with siRNAs targeting ROCK1, ROCK2, or non-targeting siRNA control (siControl). Twenty-four hours after transfection cells were plated for a clonogenic assay. Each transfection was done in triplicate, followed by clonogenic assays conducted in triplicate, and the experiments were repeated at least two times. (a) Transfection with siROCK1, but not siROCK2, resulted in significant reduction in RCC4 colony numbers in comparison to RCC4VHL. Thus, ROCK1 downregulation mimics the effect of Y-27632 treatment on viability of RCC4 cells, making it a likely target for Y-27632 causing synthetic lethality effect. Statistical analysis was performed using a paired t-test comparing numbers of colonies in each siROCK group to siControl. SEMs are shown. (b–c) The degree of each target knockdown by its specific siRNA (as indicated) was assessed by Western blot.