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. 2019 Aug;12(3):167–183. doi: 10.2174/1874467212666190304121131

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© 2019 Bentham Science Publishers

This is an open access article licensed under the terms of the Creative Commons Attribution-Non-Commercial 4.0 International Public License (CC BY-NC 4.0) (https://creativecommons.org/licenses/by-nc/4.0/legalcode), which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

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Fig. (4) — p300 (+/-) versus wild type and CBP (+/-) 3T3 cells display heightened response to canonical Wnt3a and negligible response to non-canonical Wnt5a. (a) When stimulated with Wnt3a conditioned media, 3T3 wild type (WT) and CBP (+/-) cells exhibited a small activation of the TOPFlash reporter gene construct; however, p300 (+/-) cells had a significantly larger increase in TOPFlash activity. Wnt5a alone had almost no effect on basal TOPFlash activity in the 3 cell lines. In WT and CBP (+/-) 3T3 cells, co-administration of Wnt5a and Wnt3a versus Wnt3a alone resulted in an ~20% reduction in TOPFlash activity, whereas a dramatic reduction in TOPFlash activity was observed in the p300 (+/-) cells. n ≥ 2, *p < 0.05, **p < 0.01. (b) To gain additional perspective, our studies were extended to include the survivin promoter luciferase construct pGL3-6270Luc whose activity is more CBP-dependent than the TOPFlash construct. Wnt3a caused an ~2.5-fold increase in survivin luciferase activity in 3T3 WT cells, an ~2-fold increase in 3T3 CBP (+/-) cells, and a dramatic ~50-fold increase in the p300 (+/-) cells. Wnt5a elicited a similar minimal response above control media in the WT and CBP (+/-) cells, yet had essentially no effect on the p300 (+/-) cells. Co-administration of Wnt3a and Wnt5a conditioned media led to modest reductions in survivin luciferase activity in the WT and CBP (+/-) cells compared to Wnt3a or 5a alone, whereas co-administration of Wnt3a and 5a dramatically reduced survivin luciferase activity in the p300 (+/-) cells compared to Wnt3a alone. n ≥ 2, *p < 0.05, **p < 0.01.