Journal: Advanced Science

Article Title: Aberrant Cholesterol Metabolism and Wnt/ β ‐Catenin Signaling Coalesce via Frizzled5 in Supporting Cancer Growth

doi: 10.1002/advs.202200750

Figure Lengend Snippet:

Article Snippet: L‐Wnt3a cell , ATCC , CRL‐2647.

Techniques: Staining, Western Blot, Sequencing, Control

Journal: The Journal of Biological Chemistry

Article Title: Activation of Wnt Signaling in Cortical Neurons Enhances Glucose Utilization through Glycolysis *

doi: 10.1074/jbc.M116.735373

Figure Lengend Snippet: General effects of acute Wnt3a treatment on neuronal cultures. A, treatment with Wnt3a for various lengths of time (0, 15, 30, and 90 min) did not affect the β-catenin levels. However, the treatment triggered an increase in the expression of p-GSK3β-Ser9 (inactivated form). B, using qRT-PCR, we did not observe changes in the mRNA levels of either Wnt target gene, c-jun or CaMk4, after acute treatment with Wnt3a. However, after 24 h of treatment, we observed an increase in the levels of both mRNAs, and this increase was blocked by co-incubation with Dkk1. C, using Western blot analysis, we studied the expression of the activated forms of AMPK and Akt after the acute treatment with Wnt3a and observed a strong increase in the level of the activated form of Akt after 15 and 30 min of treatment. The data represent the mean ± S.E. of n = 4 experiments, each performed in triplicate. *, p < 0.01; **, p < 0.005, Bonferroni's test.

Article Snippet: Control and Wnt3a-conditioned media were prepared from L control (ATCC: CRL-2648) and l -Wnt3a (ATCC: CRL-2647) cells.

Techniques: Expressing, Quantitative RT-PCR, Incubation, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Activation of Wnt Signaling in Cortical Neurons Enhances Glucose Utilization through Glycolysis *

doi: 10.1074/jbc.M116.735373

Figure Lengend Snippet: Effect of Wnt3a treatment on glucose uptake. A, Wnt3a treatment stimulated 2-DG uptake in a time-dependent manner. The effect of Wnt3a treatment was significant after 15 s of exposure, and the time-dependent effect was blocked by co-incubation with Dkk1. The IC50 value was similar to the values reported for GLUTs. The effect of Wnt3a was independent of both transcription and synaptic effects of Wnt3a and was blocked by co-treatment with Dkk1, AZD5363, and TCS183. B, the initial uptake of trace amounts of 2-DG (at 15 s) was measured in the presence of increasing concentrations of unlabeled glucose (0–30 mm) under the control conditions and following Wnt3a treatment (15 min). The data represent the mean ± S.E. of n = 5 experiments, each performed in triplicate. The continuous lines in each plot correspond to the best non-linear regression fit of a rectangular hyperbola to the data using SigmaPlot 12 software.

Article Snippet: Control and Wnt3a-conditioned media were prepared from L control (ATCC: CRL-2648) and l -Wnt3a (ATCC: CRL-2647) cells.

Techniques: Incubation, Control, Software

Journal: The Journal of Biological Chemistry

Article Title: Activation of Wnt Signaling in Cortical Neurons Enhances Glucose Utilization through Glycolysis *

doi: 10.1074/jbc.M116.735373

Figure Lengend Snippet: Effects of Wnt3a on GLUT3 expression, HK activity, and the glycolytic rate. A, using Western blot analysis, we observed that Wnt3a treatments for various lengths of time did not affect GLUT3 expression. B, Wnt3a treatment increased HK activity. This effect was blocked by Dkk1, TCS183, and AZD5363. C, Wnt3a treatment increased the glycolytic rate in a time-dependent manner, and this effect was blocked by co-incubation with Dkk1. The glycolytic rate was also increased by other canonical Wnt activators, such as LiCl. The Wnt3a-induced increase in the glycolytic rate was independent of the synaptic effects of Wnt3a and dependent on Akt activation. D, using Western blot analysis, we studied the expression of the activated form of PFKFB2, and we observed an increase in the level of this form after Wnt3a treatment. The data represent the mean ± S.E. of n = 5 experiments (4 for D), each performed in triplicate. *, p < 0.01; **, p < 0.005, Bonferroni's test.

Article Snippet: Control and Wnt3a-conditioned media were prepared from L control (ATCC: CRL-2648) and l -Wnt3a (ATCC: CRL-2647) cells.

Techniques: Expressing, Activity Assay, Western Blot, Incubation, Activation Assay

Journal: The Journal of Biological Chemistry

Article Title: Activation of Wnt Signaling in Cortical Neurons Enhances Glucose Utilization through Glycolysis *

doi: 10.1074/jbc.M116.735373

Figure Lengend Snippet: Activation of the Wnt pathways does not affect PPP activity. A, treatment with Wnt3a did not affect the activity or the levels of the G6PDH enzyme. B, the oxidation of glucose through the PPP did not change following treatment with Wnt3a ligand. C, the increase in the glycolytic rate was correlated with an increase in the generation of ADP, without changes in PPP activity. The data represent the mean ± S.E. of n = 5 experiments, each performed in triplicate. *, p < 0.01; **, p < 0.005, Bonferroni's test.

Article Snippet: Control and Wnt3a-conditioned media were prepared from L control (ATCC: CRL-2648) and l -Wnt3a (ATCC: CRL-2647) cells.

Techniques: Activation Assay, Activity Assay

Journal: The Journal of Biological Chemistry

Article Title: Activation of Wnt Signaling in Cortical Neurons Enhances Glucose Utilization through Glycolysis *

doi: 10.1074/jbc.M116.735373

Figure Lengend Snippet: Wnt pathway activation stimulates 2-DG uptake in hippocampal slices. A, in slices, Wnt3a treatment induced a time-dependent increase in the intracellular accumulation of 2-DG. This effect was blocked by Dkk1 and TCS-183. B, Wnt3a treatment increased HK activity. C, we did not observe changes in the activity of G6PDH in hippocampal slices after Wnt3a treatment. D, Wnt3a treatment induced a large increase in the glycolytic rate. E, treatment with Wnt3 did not affect the activity of the PPP in hippocampal slices. The data represent the mean ± S.E. of n = 5 experiments, each performed in triplicate. *, p < 0.01; **, p < 0.005, Bonferroni's test.

Article Snippet: Control and Wnt3a-conditioned media were prepared from L control (ATCC: CRL-2648) and l -Wnt3a (ATCC: CRL-2647) cells.

Techniques: Activation Assay, Activity Assay

Journal: The Journal of Biological Chemistry

Article Title: Activation of Wnt Signaling in Cortical Neurons Enhances Glucose Utilization through Glycolysis *

doi: 10.1074/jbc.M116.735373

Figure Lengend Snippet: Schematic representation of the effect of Wnt3a on glucose metabolism in cortical neurons. Wnt3a stimulates glucose uptake and HK activity. Downstream, Wnt3a increases the glycolytic rate. The increase in the glycolytic rate is correlated with an increase in the generation of ADP, without changes in PPP activity. The effects of Wnt3a over the glucose metabolism was independent of the synaptic effect of Wnt3a, because the co-incubation both d-APV and CNQX does not block the metabolic effect. The mechanism of action of Wnt3a is dependent of activation of the metabolic sensor Akt, suggesting a complex network between the metabolic pathways.

Article Snippet: Control and Wnt3a-conditioned media were prepared from L control (ATCC: CRL-2648) and l -Wnt3a (ATCC: CRL-2647) cells.

Techniques: Activity Assay, Incubation, Blocking Assay, Activation Assay