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. 2020 May 21;5(10):e133365. doi: 10.1172/jci.insight.133365

Figure 6. Persistent DNA damage–induced NLRP12 maintains the function of HSCs from patients with FA and aged donors.

Figure 6

(A) Increased NLRP12 expression in HSPCs from patients with FA or aged donors under DNA damage. Real-time qPCR mRNA measurement of NLRP12 in BM CD34+ cells from healthy donors (Normal) and patients with FA (upper) or young and aged donors (lower) after 16-hour culture in the presence or absence of MMC (10 nM). Samples were normalized to the level of GAPDH mRNA. Results are shown as means ± SD of 3 independent experiments (n = 6–9 per group). (B) Schematic presentation of experimental design. (C and D) Persistent DNA damage–induced NLRP12 maintains the function of HSCs from patients with FA or aged donors. BM CD34+ cells from the indicated donors or patients were transduced with scramble shRNA or shRNA targeting NLRP12. Two thousand sorted GFP+ cells were transplanted into sublethally irradiated NSGS recipients followed by MMC injection at 2 weeks posttransplant. Human engraftment was analyzed 16 weeks after transplant by flow cytometry using anti-human CD45 antibody. Results are shown as means ± SD of 3 independent experiments (n = 9 per group). *P < 0.05; **P < 0.01; ***P < 0.001. Paired or unpaired 2-tailed Student’s t test was used for 2-group comparison and 1-way ANOVA for comparison of more than 2 groups.