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. 2023 Dec 4;14(12):791. doi: 10.1038/s41419-023-06274-1

Fig. 3. SOX2 decreases the enrichment of H3K27 methylation and transcriptionally activates AC005392.2 expression.

Fig. 3

a HCT116 cells were transfected for 72 h with a SOX2 clone and subjected to actinomycin D (2.5 μg/ml) treatment for the indicated times. qRT-PCR was performed to analyze the expression of AC005392.2 (mean ± SEM; n = 3). b A schematic map of potential SOX2-binding sites in the promoter of AC005392.2, shown according to the JASPAR database. c A ChIP assay was performed to confirm the interaction between the SOX2 and AC005392.2 promoter in HCT116 and SW620 cells after transfection for 72 h with SOX2 clones or SOX2 shRNA. An IgG antibody was used as a negative control (mean ± SD; n = 3, two-tailed Student’s t test). d AC005392.2 promoter-driven luciferase activity was assessed in HCT116 and SW620 cells transfected with a SOX2 clone or SOX2 shRNA using a luciferase reporter assay (mean ± SD; n = 3, two-tailed Student’s t test). e, f Luciferase activity of the AC005392.2 promoter was examined in HCT116 cells and SW620 cells transfected with a truncated AC005392.2 promoter (e) or deletion mutant of the AC005392.2 promoter (f, mean ± SD; n = 3, two-tailed Student’s t test). g Western blotting examining the expression of histone methylation and acetylation modification in SOX2-overexpressing HCT116 and SOX2-depleted SW620 cells. h A ChIP assay identifying the expression of H3K27me3 at the promoter of AC005392.2 in SOX2-overexpressing HCT116 and SOX2-depletion SW620 cells (mean ± SD; n = 3, two-tailed Student’s t test). ***p < 0.001, and ****p < 0.0001.