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. 2016 May 29;173(14):2237–2249. doi: 10.1111/bph.13503

Figure 5.

Figure 5

Mdk‐Ab treatment diminished the negative influence of Mdk on β‐catenin signalling in preosteoblastic cells. (A) Alpl gene expression in MC3T3‐E1 cells on day 5 of differentiation after 6 h of treatment with Mdk and the Mdk‐Ab. B2M was used as the housekeeping gene, and gene expression values were normalized to the pre‐differentiation values (dotted line). n = 6. (B) Alpl protein expression in MC3T3‐E1 cells on day 5 after 6 h of stimulation. α‐Tubulin was used as control. n = 3. (C) Acan gene expression in ATDC5 cells on day 5 of differentiation after 6 h of stimulation with Mdk and the Mdk‐Ab. B2M was used as the housekeeping gene, and gene expression values were normalized to the unstimulated control (dotted line). (D) ATDC5 and MC3T3‐E1 cells were incubated without or with recombinant Mdk for 1 h, and immunoprecipitation was performed with Mdk‐Ab. n = 6. (E) Phospho‐LRP‐6, LRP‐6, β‐catenin and active β‐catenin protein expression in MC3T3‐E1 cells after 6 h of incubation. n = 3. (F) Lef1 gene expression in MC3T3‐E1 cells on day 5 of differentiation after 6 h of treatment with Mdk and the Mdk‐Ab. B2M was used as the housekeeping gene, and gene expression values were normalized to the pre‐differentiation values (dotted line). n = 6.