Figure 6.

Knocking down CRL4B^DCAF11 complex components inhibits osteosarcoma cell growth. (A) Knocking down CRL4B^DCAF11 complex components leads to the accumulation of p21. U2OS cells transfected with Con-shRNA, CUL4B-shRNA, DDB1-shRNA, or DCAF11-shRNA. After 48 h incubation, cells were lysed and probed with anti-CUL4B, DDB1, DCAF11, p21, CDK2, or β-Actin antibody. (B) Knocking down CRL4B^DCAF11 complex components inhibits osteosarcoma cell proliferation. Cells used in A were applied to the MTT assay to evaluate cell proliferation, and cell viability was determined at 490 nm. (C) Knocking down CRL4B^DCAF11 complex components induces cell cycle arrest at S phase. Cells used in A were subjected to flow cytometry analysis of cell cycle distribution. (D,E) Knocking down CRL4B^DCAF11 complex components decreased colony formation rates. Cells (1 × 10³) used in A were seeded onto 12-well plates, and cells were cultured with 0.1 ml of fresh medium containing 0.5% FBS for two weeks. Then, cells were stained with 0.5% crystal violet (D), and colony numbers were counted by a gel documentation system (E).