Fig. 2

ASL expression alters growth in normal kidney and ccRCC cells. A mRNA expression levels of ASL in HK-2 cells post-knockdown with two independent shRNAs. Scrambled shRNA was used as a control. B Protein levels of ASL in HK-2 cells after shRNA knockdown. Actin is the loading control. C, D ASL knockdown enhances HK-2 cell growth in C 2D and D 3D soft agar colony-forming assay. In C, cells were grown in 1% FBS and standard DMEM. Error bars represent SEM from three replicate wells. **p < 0.01, *p < 0.05. E Ectopic expression of ASL in 769-P ccRCC polyclonal populations. HK-2 cells were used as physiologic control. HSP90 and Actin serve as loading controls. F–H ASL re-expression in 769-P cells suppresses growth in F 2D, G 3D matrigel spheroids, and H 3D soft agar colony-forming assay. 769-P cells expressing empty vector were used as a control. In F, cells were grown in 1% FBS and standard DMEM. **p < 0.01, *p < 0.05