Figure 7: PKD1 impaired osteoblast differentiation through JAK1/STAT3 and p38 signaling pathway.
(a) After days 3, 5 differentiating, MC3T3-E1 cells were treated with CRT0066101 or vehicle for 24 hours and then underwent immunoblotting to detect PKD1, PKD1 phosphorylation activity, JAK1, STAT3 and phosphorylation (ser727). Photoshop analyzed the density of bands on western blot. For each marker band, values were normalized to control and the density of control was set at 1. (b) Western Blot analysis of differentiating primary calvarial preosteoblasts showed decreased expression of p38,P-p38,STAT3,P-STAT3-ser727, y705 and PCNA in KO cells (0, 12th day). n = 3, WT or KO mouse. (c) MC3T3-EI cells were transfected with si-PKD1 and negative control (si-CTL), pcDNA PKD1 and empty vector plasmid for 60 hours to analyze osteoblast regulators (Runx2, OPN,OSX and OCN) and relative pathway markers (STAT3 and p38). (d) MC3T3-EI cells were transfected with si-STAT3 and negative control (si-CTL) for 60 hours. Uncropped western blot images corresponding to Fig. 7(a,b,c and d) were shown in Supplementary Information file.
