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. Author manuscript; available in PMC: 2023 Apr 14.
Published in final edited form as: Clin Cancer Res. 2022 Oct 14;28(20):4466–4478. doi: 10.1158/1078-0432.CCR-22-0384

Figure 3. The PHE uniquely controls HOXD13 expression in Ewing sarcoma.

Figure 3.

All cells express dCas9-KRAB-mcherry and sgRNAs targeting either a negative control region (neg), the SOX2 GGAA enhancer, or PHE (1-3). DNA was isolated 8 days after sgRNA transduction. A) ChIP-qPCR for H3K9me3, B) EWS::FLI1, and C) H3K27ac at the PHE region. qRT-PCR of HOXD13 levels in D) A673 and E) CHLA10 cells. F) HiChIP looping data depicting the HOXD13 promoter:enhancer interactions in the C-DOM region in A673 cells (14). G) Model of EWS::FLI1 regulation of HOXD13 in Ewing sarcoma cells (biorender). Error bars are representative of SEM from at least three independent experiments. * p<0.05; ** p<0.01; Two tailed t-test; Two-way ANOVA; Sidak’s multiple comparison test.