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. 2022 Oct 10;25(4):635–647. doi: 10.1093/neuonc/noac231

Figure 4.

Figure 4.

HOXA3 activates the transcription of KDM6A and modifies H3K27me3 of HK2 and PKM2 promoter. (A) Gene set enrichment assay of IlluminaHiSeq (n = 172) the cancer genome atlas (TCGA) showed significant enrichment of gene sets involved in the methylation with the abundant genes being upregulated (cutoff: median). (B) ChIP-qPCR analysis of H3K4me3, H3K9me3, and H3K27me3 at HK2 and PKM2 promoter in GBM cells expressing shCtrl or shHOXA3. (C) Immunoblotting of HOXA3, EZH2, KDM6A, and H3K27me3 in the indicated GBM cells expressing shCtrl, shHOXA3, or shHOXA3/HOXA3. (D) Predicted sequences of HOXA motif in KDM6A promoter by JASPAR. (E) ChIP-qPCR analysis of HOXA3 at the indicated KDM6A promoter in GBM cells expressing shCtrl or shHOXA3. (F) The KDM6A promoter-reporter constructs were co-transfected with shCtrl or shHOXA3 vector into HEK293T cells. The promoter activities were presented. (G) Immunoblotting of HOXA3, KDM6A, HK2, and PKM2 in the indicated GBM cells expressing shCtrl, shHOXA3, or shHOXA3/KDM6A. (H) ChIP-qPCR analysis of H3K27me3 at HK2 and PKM2 promoter in GBM cells expressing shCtrl, shHOXA3, or shHOXA3/KDM6A. All data were shown as means and SD (n = 3), **P < .01.