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. 2022 Feb 8;7(3):e153768. doi: 10.1172/jci.insight.153768

Figure 4. KDM5D loss promotes tumorigenesis and increased DNA damage.

Figure 4

(A) Venn diagram showing the enrichment of the significantly downregulated genes in mLOY AML patients (top 200) and AML mice. (B and C) The expression levels of KDM5D in mLOY AML patients with AML1-ETO (B, n = 5; C, n = 3) and other AML1-ETO+ patients (B, n = 4; C, n = 3) in 2 AML cohorts (TARGET AML and BEAT AML), from analysis of The Cancer Genome Atlas data. *P < 0.05, **P < 0.01 (2-tailed t test). (D) Kaplan-Meier tumor-free survival curve of recipient mice with sgScr; AML1-ETO and sgKdm5d; AML1-ETO AML cells (n = 6). *P < 0.05 (log-rank test). (E) Left: Representative immunofluorescence images of γH2AX foci in sgScr and sgKdm5d HSPCs. Scale bars: 10 μm. Red, γH2AX; blue, DAPI-labeled DNA. Right: Plot of γH2AX foci per cell, shown as the mean ± SD. ****P < 0.0001 (2-tailed Mann-Whitney test). (F) Left: Representative images of comet assay of sgScr and sgKdm5d HSPCs. Scale bars: 50 μm. Right: Plot of comet assay of sgScr and sgKdm5d HSPCs. The tail moment is shown as the mean ± SD. ****P < 0.0001 (2-tailed Mann-Whitney test). (G) Left: Representative images of comet assay of sgSsty1-sgCas9 HSPCs with truncated-Kdm5d overexpression (bottom, the truncated KDM5D is a 659–amino acid protein consisting of aa 1–695) and vector only (top) was used as a negative control. Scale bars: 50 μm. Right: Plot of comet assay of sgSsty1-sgCas9 HSPCs with truncated-Kdm5d overexpression and vector only. The tail moment is shown as the mean ± SD. ****P < 0.0001 (2-tailed Mann-Whitney test). (H) Left: Representative images of comet assay of sgSsty2-sgCas9 HSPCs with truncated-Kdm5d overexpression (bottom) and vector only (top). Scale bars: 50 μm. Right: Plot of comet assay of sgSsty2-sgCas9 HSPCs with truncated-Kdm5d overexpression and vector only. The tail moment is shown as the mean ± SD. ****P < 0.0001 (2-tailed Mann-Whitney test).