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. Author manuscript; available in PMC: 2021 May 17.
Published in final edited form as: J Cell Physiol. 2019 Dec 23;235(6):5293–5304. doi: 10.1002/jcp.29415

FIGURE 4.

FIGURE 4

Inhibition of osteogenic differentiation by +JQ1 is reversible in MC3T3 cells. Subconfluent MC3T3 cells were treated from Day 0 to Day 3 with 100 nM +JQ1. Osteogenic differentiation supplements were added on Day 0 and replenished throughout the entire culture period. (a) Quantitative reverse transcription polymerase chain reaction (RT-qPCR) performed on messenger RNA (mRNA) isolated on Day 3 (n = 3, mean ± STD). (b) Alkaline phosphatase (Alpl) enzymatic activity measured on Day 6 (n = 3, mean ± STD). (c) mRNA expression levels assessed via RT-qPCR for indicated genes throughout the osteogenic differentiation time course (n = 3, mean ± STD). (d) Representative image (left) and ImageJ quantification (right) of alizarin red staining for hydroxyapatite deposition. All error bars represent ± STD of three biological replicates. Statistical significance to vehicle (DMSO) control group indicated as follows: ^p < .05, *p < .01, #p < .001, n.s.p > .05