Knockdown of Brd2 and/or Brd4 impairs osteogenic differentiation of MC3T3 cells. (a) Western blot analysis of protein lysates collected 2 days after small interfering RNA (siRNA) transfection. (b) Quantitative reverse transcription polymerase chain reaction (RT-qPCR) analysis performed on messenger RNA (mRNA) isolated 4 days post-transfection (n = 3, mean ± STD). (c) Pico Sirius Red staining and quantification for collagen deposition performed 7 days after induction of osteogenic differentiation (n = 3, mean ± STD). (d) Alkaline phosphatase (Alpl) enzymatic activity measured on Day 14 of osteogenic differentiation (n = 3, mean ± STD). (e) RT-qPCR conducted on mRNA isolated 3 days after induction of osteogenic differentiation (n = 3, mean ± STD). All error bars represent ± STD of three biological replicates. Statistical significance to nontargeting control group indicated as follows: ^p < .05, *p < .01, #p < .001