Figure 7.
ATF2, specifically phosphorylated ATF2 at Thr69/71, is a promoter of DEK transcription. (A) The list shows the most potential transcription factors (TFs) regulating DEK and their motif using the ChIPBase database. (B) Gene correlation analysis between ATF2 and DEK in human whole blood cells (GTEx database) was conducted using GEPIA server. Correlation coefficient (r) and P value (r = 0.9, P = 0) were calculated using Pearson’s correlation analysis. (C) Representative heatmaps and line plots show p-ATF2 (Thr69/71) and p-ATF2 (Ser490/498) CUT&Tag peaks ± 5 kb from TSS in HSCs (CD48−CD150+LSK) from WT C57BL/6J mice. (D) Statistics and quantification of CUT&Tag signal in HSCs (n = 2 samples per group). (E) Distribution of p-ATF2 (Thr69/71) and p-ATF2 (Ser490/498) CUT&Tag peaks across the indicated gene loci in HSCs. (F) Violin plots show distribution of CUT&Tag signal at genomic regions of indicated genes in HSCs. (G) Presentation of partial amino acid sequence of human and mouse ATF2 and the design of ATF2 mutations (indicated with red font). (H) The bar graph shows the luciferase activity under indicated conditions (n = 5 samples per group). (I) qRT-PCR analysis and the bar graph shows the expression of ATF2 and DEK gene in transduced HSCs (n = 4 samples per group). (J) The bar graphs show the MFI of DEK in transduced HSCs using flow cytometry staining and analysis (n = 5 samples per group). (K) Representative flow plots show p-ATR staining in transduced HSCs. The right bar graphs show the MFI of p-ATR and p-CHK1 in HSCs (n = 5 samples per group). (L) Representative flow plots show the EdU+ in transduced HSCs. The right bar graph shows the frequency of EdU+ cells in HSCs (n = 5 samples per group). The data presented in panels C–L are representative of two independent experiments. The results shown in panels D and F are illustrated as violin plots, and other panels are shown as mean ± SD. Statistical analyses were performed using a two-tailed unpaired Student’s t test for panels D and F and one-way ANOVA followed by Tukey’s post hoc test for panels H–L. P values are presented as follows: *P < 0.05, **P < 0.01, and ***P < 0.001; n.s., not significant. Refer to the image caption for details.

ATF2, specifically phosphorylated ATF2 at Thr69/71, is a promoter of DEK transcription. (A) The list shows the most potential transcription factors (TFs) regulating DEK and their motif using the ChIPBase database. (B) Gene correlation analysis between ATF2 and DEK in human whole blood cells (GTEx database) was conducted using GEPIA server. Correlation coefficient (r) and P value (r = 0.9, P = 0) were calculated using Pearson’s correlation analysis. (C) Representative heatmaps and line plots show p-ATF2 (Thr69/71) and p-ATF2 (Ser490/498) CUT&Tag peaks ± 5 kb from TSS in HSCs (CD48−CD150+LSK) from WT C57BL/6J mice. (D) Statistics and quantification of CUT&Tag signal in HSCs (n = 2 samples per group). (E) Distribution of p-ATF2 (Thr69/71) and p-ATF2 (Ser490/498) CUT&Tag peaks across the indicated gene loci in HSCs. (F) Violin plots show distribution of CUT&Tag signal at genomic regions of indicated genes in HSCs. (G) Presentation of partial amino acid sequence of human and mouse ATF2 and the design of ATF2 mutations (indicated with red font). (H) The bar graph shows the luciferase activity under indicated conditions (n = 5 samples per group). (I) qRT-PCR analysis and the bar graph shows the expression of ATF2 and DEK gene in transduced HSCs (n = 4 samples per group). (J) The bar graphs show the MFI of DEK in transduced HSCs using flow cytometry staining and analysis (n = 5 samples per group). (K) Representative flow plots show p-ATR staining in transduced HSCs. The right bar graphs show the MFI of p-ATR and p-CHK1 in HSCs (n = 5 samples per group). (L) Representative flow plots show the EdU+ in transduced HSCs. The right bar graph shows the frequency of EdU+ cells in HSCs (n = 5 samples per group). The data presented in panels C–L are representative of two independent experiments. The results shown in panels D and F are illustrated as violin plots, and other panels are shown as mean ± SD. Statistical analyses were performed using a two-tailed unpaired Student’s t test for panels D and F and one-way ANOVA followed by Tukey’s post hoc test for panels H–L. P values are presented as follows: *P < 0.05, **P < 0.01, and ***P < 0.001; n.s., not significant.

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