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. 2015 Apr 23;33(5):1419–1433. doi: 10.1002/stem.1944

Figure 7.

Figure 7

β‐Catenin‐induced Slug upregulation reinforces human embryonic stem cell (hESC) differentiation by downregulating E‐cadherin and upregulating Brachyury. (A, B): Long‐term but not short‐term BIO treatment enhances Slug expression in hESCs (A, Q‐PCR; B, Western blot). (C): hESCs cotransfected with EcadΔβ and βS33Y, but not with vectors (pcDNA) or with wild‐type E‐cadherin (WT‐Ecad) and βS33Y, enhances Slug expression (Q‐PCR). (D): Over‐expression of EcadΔβ together with short‐term Wnt3a treatment (100 ng/ml) promotes Slug expression in hESCs (Q‐PCR). (E): hESCs infected with pLKO‐siSlug plasmid (siSlug) for 2 days reduce Slug expression (Western blot). Scrambled: pLKO‐puro vector with a scrambled sequence that does not target any mRNA. (F–H): Slug knockdown (pre‐siSlug) followed by BIO treatment for 4 days results in a decrease of differentiation marker Brachyury (F, Q‐PCR; G, Western blot) and clonogenic capacity (H, clonogenic assays, n = 3) in hESCs. All data are mean ± SD, *, p <.05; **, p < .01. (I): The proposed working model (detailed in Supporting Information Fig. S7).