Silencing of Gli1 increasesmineralization inMC3T3preosteoblast in the absence or presence of EPZ6438. Diagram of the experimental protocol for treatment of MC3T3 with siCtrl or siGli1 and with vehicle (Veh) or 2.5 μM EPZ6438 shown in panels B–D (A). Gli1 mRNA levels as evaluated by RT-qPCR on day 2 (B). RT-qPCR analysis for bone-related markers relative to Gapdh (set at 100) on day 9 (C). Alizarin red staining on day 43 (D). All error bars represent ±SD of three experimental replicates. (ns = not significant, ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001, and ∗∗∗∗p ≤ 0.0001). RT-qPCR, real-time quantitative PCR.