Fig. 4 Loss of Zic5 derepresses GLUT1/SLC2A1 gene expression.

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Fig. 5 Correlation of RNA expression and protein abundance.

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Fig. 4 Loss of Zic5 derepresses GLUT1/SLC2A1 gene expression. Loss of Zic5 derepresses GLUT1/SLC2A1 gene expression. (A) Glycolysis-related gene expression change in Zic5 WT and KO cells are shown in the color-coded format. The ratio indicates the log fold change (logFC). (B) Loss of Zic5 increased GLUT1/SLC2A1 mRNA level as shown by RNA-seq. (C) Zic5 binding at the promoter of GLUT1/SLC2A1 gene is lost in Zic5 KO clones. (D) Quantitative reverse transcription polymerase chain reaction (qRT-PCR) showing that overexpression of Zic5 but not Zic5-N decreases GLUT1/SLC2A1 mRNA level. (E) qRT-PCR showing that the KO of Zic5 in 293T cells increases the GLUT1/SLC2A1 mRNA level. (F) GLUT1 protein levels in Zic5 WT and KO HCT116 cells are measured by flow cytometry. (G) Western blot showing GLUT1 expression levels in Zic5 WT and KO cells. (H) Glucose uptake in Zic5 WT and KO HCT116 cells is measured by the amount of 2-NBDG taken up by the cells. The fluorescent intensity is quantified by the plate reader (n = 4). The pairwise Wilcoxon rank sum tests were used to calculate the significance of C16 and C18 cells glucose uptake compared with WT cells. *P < 0.05. (I) Colony formation assay showing the effect of GLUT1 depletion in Zic5 WT and KO cells. Zibo Zhao et al. Sci Adv 2019;5:eaax0698 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).