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Figure 1 | Molecular Cancer

Figure 1

From: Aldolase positively regulates of the canonical Wnt signaling pathway

Figure 1

Aldolase isomers activate the canonical Wnt signaling pathway. (A) HEK293T cells were co-transfected with GFP-ALDOC, GFP-ALDOB, an empty vector, pTOPFLASH or pFOPFLASH and β-gal. Forty eight hours later, cells were harvested and lysates were measured for their luciferase activity, (upper panel) and levels of endogenous active β-catenin (lower panel). Anti-GFP antibody was used to detect Aldolase proteins. Data presented describe relative mean values and standard deviations of four independent experiments performed in duplicates. Asterisks indicate significant difference in Wnt signaling levels (** p < 0.01, Student’s t test). (B) HEK293T cells were transfected with different concentrations of siALDOC RNA. Seventy-two hours later cells were harvested and levels of endogenous ALDOC were determined (upper panel). Alternatively, HEK293T cells were co-transfected with GFP-β-catenin or an empty vector along with the pTOPFLASH or pFOPFLASH and β-gal, and 24 h later the cells were transfected with siALDOC (100 mM). Forty eight hours later the cells lysates were measured for their luciferase activity (lower panel). Asterisks indicate significant difference in Wnt signaling levels (* p < 0.05, ** p < 0.01, Student’s t test). (C) HEK293T or HeLa cells were transfected with the two Aldolase isomers as indicated. Forty eight hours later, lysates were subjected to western blot analysis using specific antibodies to distinct members of the Wnt signaling pathway. An anti-GFP antibody was used to detect Aldolase. Asterisk points to a non-specific band. (D) HEK293T cells were co-transfected with GFP-ALDOA, GFP-ALDOB or an empty vector, pTOPFLASH or pFOPFLASH and β-gal. Forty eight hours later, cells were harvested and lysates were measured for their luciferase activity, (upper panel) and levels of endogenous active β-catenin (lower panel). Data presented describe relative mean values and standard deviations of 3 independent experiments performed in duplicates or triplicates. Asterisks indicate significant difference in Wnt signaling levels (** p < 0.01, Student’s t test). Tubulin served as a loading control in all experiments.

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