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Fig. 5 | Biology Direct

Fig. 5

From: Dynamic expression of IGFBP3 modulate dual actions of mineralization micro-environment during tooth development via Wnt/beta-catenin signaling pathway

Fig. 5

Results demonstrating IGFBP3 regulate Wnt signaling pathway via DKK1. A RT-qPCR compared tooth development related markers, including DSPP (a1) and DMP1 (a2), and osteogenic markers, including OSX (a3), OPN (a4), OCN (a5) and ALP (a6) between lv5 hDPSCs, IGFBP3-over hDPSCs and IGFBP3-over hDPSCs treated with DKK1 inhibitor WAY-262611 (0.1 μM and 1 μM) for 14 days. Western blot analysis compared protein levels of DSPP, OSX, OPN, OCN and ALP between these groups (b). Alizarin red staining compared mineralized deposition between these groups after 14 days osteogenic induction, scale bar: 200 μm (c). B Location specific western blot analysis compared cytoplasmic beta-catenin (CTNNB1) levels and nuclear CTNNB1 levels between lv5 hDPSCs and IGFBP3-over hDPSCs (a), with quantitative analysis of nuclear CTNNB1 (b). Cyt: cytoplasmic, Nuc: nuclear, LAMIN B1 serve as the loading control for nuclear protein, β-ACTIN serve as the loading control for total protein. C Immunofluorescence analysis comparing CTNNB1 transnucleation by immunofluorescence. Scale bar: 50 μm. (E) Coimmunoprecipitation showed that anti-IGFBP3 pulled down DKK1 protein. n = 3, *p < 0.05, **p < 0.01, ***p < 0.001

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