Fig. 12.

shRNA-mediated silencing of PKD2 causes G2/M cell cycle arrest in bladder cancer cells. UMUC1 cells were transduced with lentivirus coding for human shRNA sequences for either PKD2 (shPKD2#1 and shPKD2#2), PKD3 (shPKD3#1 and shPKD3#2), or the control (shNTC). Cells were harvested after transduction and stained with propidium iodide. The cell cycle distribution of the propidium iodide-labelled cells was analyzed by flow cytometry, as described in “Materials and methods”. The percentage of cells in each phase of the cell cycle was calculated using FlowJo v14.0 and is indicated in each cell cycle profile (a). The assay was performed twice and one profile for each treatment is shown in (a). The average numeric data of the two biological replicates for both PKD2-depleted and PKD3-depleted UMUC1 cells are presented in b. Note the accumulation of the cells in the G2/M phase in PKD2-knockdown UMUC1 cells, but not in PKD3-knockdown UMUC1 cells (b). shRNA short hairpin RNA