ZFIN Image: Zhang et al., 2020, Fig. 3

Image

Figure Caption

Fig. 3 Figure 3. Overexpression or knockdown of VBP1 promotes protein degradation of TCF/LEFs. A, exogenous Tcf/Lef protein levels in control or VBP1-overexpressing HEK293T cells. Similar results were obtained from four experiments. B, endogenous TCF7L2 protein level in control or VBP1-overexpressing HEK293T cells. Similar results were obtained from three experiments. C, changes in exogenous TCF7L2 protein level in VBP1-overexpressing HEK293T cells treated with MG132. Cells transfected with indicated plasmid DNA and treated with or without 10 µm MG132 for 8 h. Similar results were obtained from three experiments. D, TCF7L1 and TCF7L2 protein levels in control or VBP1-knockdown HEK293T cells. Similar results were obtained from three experiments. E, qRT-PCR analysis of transcriptional levels of TCF7L1 and TCF7L2 in control and VBP1-knockdown HEK293T cells. Data are from three independent experiments with individual data points shown. F, representative image of TCF7L2 protein levels in control or VBP1-knockdown HEK293T cells treated with CHX in a time series. G, quantitation of relative protein levels from the two isoforms of TCF7L2, TCF7L2E (left panel) and TCF7L2M/S (right panel), in (F), respectively. The band intensity was quantified using ImageJ software. Data are from three independent experiments. H, TCF7L1 and TCF7L2 protein levels in control or VBP1-knockdown HCT116 cells. Similar results were obtained from three experiments. I, changes in TCF7L2 protein levels in VBP1-knockdown HEK293T cells treated with MG132. VBP1-depleted HEK293T cells treated with or without 10 µm MG132 for 8 h. Similar results were obtained from three experiments. J, changes in TCF7L2 protein levels in VBP1-knockdown HCT116 cells treated with MG132. VBP1-knockdown HCT116 cells treated with or without 10 µm MG132 for 8 h. Similar results were obtained from three experiments. K, ubiquitylation assays in HEK293T cells transfected with indicated plasmids. HIF-1α was used as a positive control of ubiquitylation promoted by VBP1. Similar results were obtained from three experiments. The values are mean ± S.D. Unpaired t test. *p < 0.05; **p < 0.01; ***p < 0.001; ns, nonsignificant.

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ J. Biol. Chem.