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. 2013 Dec 9;8(12):e81348. doi: 10.1371/journal.pone.0081348

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© 2013 Kanojia et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Western blot depicts ablation of SPAG9 protein in UM-UC-3 cells transfected with SPAG9 siRNA. β-Actin was used as a control. (B) Knockdown of SPAG9 by siRNA inhibited cell growth in UM-UC-3 cells in vitro determined by MTT assay. (C) Silencing of SPAG9 expression arrested cell cycle progression in G₀–G₁ phase in SPAG9 siRNA transfected cells as compared with only cells or scrambled siRNA transfected cells. (D) Histogram analysis confirmed that in SPAG9 siRNA transfected cells accumulated in G₀–G₁ phase. (E) Western blot analysis of cell cycle molecules in SPAG9 ablated UM-UC-3 cells depicted up-regulation of CDK inhibitors including p21 and p16. The up-regulation of p21 and p16 was preceded by down-regulation of cyclin E, cyclin D, cyclin B, CDK4 and CDK1. β-Actin was used as an internal loading control. [M1- G₀–G₁ phase_; M2- S phase and M3- G₂-M phase].