Fig. 5

Wnt/β-catenin signaling regulates HySp5 expression. a HySp5 expression in intact Hydra exposed to ALP for two days, detected by WISH (left) (3 independent experiments) and qPCR (right). Each point represents an independent replicate. Scale bar: 250 μm. b Map of the 2’992 bp genomic region encompassing the Sp5 promoter from the Hm-105 strain and phylogenetic footprinting plot comparing this region in Hm-105, H. oligactis and H. viridissima. Pink peaks and binding sites as in Fig. 4. Blue rectangles: regions tested for Sp5 binding in ChIP-qPCR assays. c HySp5 promoter activity measured in HEK293T cells co-transfected with HySp5–2992:Luc and plasmids expressing activators of Wnt/β-catenin signaling, human WNT3, LRP6, Δβ-Catenin. d ChIP-qPCR assays performed in HEK293T cells expressing HySp5–2992:Luc together with HySp5–420 or HySp5-ΔDBD. Source Data are provided as a Source Data file. e HySp5 promoter activity measured in HEK293T cells expressing HySp5–2992:Luc with huΔβ-Catenin, HySp5–420 or HySp5-ΔDBD. f Immunoprecipitation (IP) of HA-tagged HySp5–420 expressed in HEK293T cells together or not with huΔβ-Catenin (upper) or huTCF1 (lower). IP was performed with an anti-HA antibody and Co-IP products were detected with the anti-β-catenin or anti-TCF1 antibodies. Same results were obtained in two independent experiments. Each data point in (c-e) represents one biological independent experiment. Statistical p values: *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001 (unpaired t test). Error bars indicate SD. PP primer pair, RLA relative luciferase activity