Fig. 5: Palbociclib prevents genotoxicity in HSPCs. | Nature Communications

Fig. 5: Palbociclib prevents genotoxicity in HSPCs.

From: Single-cell multiplex approaches deeply map ON-target CRISPR-genotoxicity and reveal its mitigation by palbociclib and long-term engraftment

Fig. 5: Palbociclib prevents genotoxicity in HSPCs.

a Experimental design. HSPCs from CB were edited with Cas9 RNP and gRNA targeting HBG1/2 promoters with or without palbociclib exposure (1 µM, overnight before transfection and for 24 h or 48 h post-transfection). Cell cycle analysis, nuclear abnormalities, and micronuclei count (day 2), Nanopore sequencing, and scSNP-DNAseq (day 4) were performed. b Histogram of cell cycle after 12 h and 36 h of cell culture with or without palbociclib (Mean ± SD, n = 3 independent experiments), and after 48 h of clearance. c Left panel, fluorescence microscopy: % of nuclear abnormalities quantified by DAPI staining at day 2 (Mean ± SD, n = 3 independent experiments, with 300, 100, and 100 cells, respectively. Paired t-test (two-tailed)). Right panel, % of micronuclei (MN) quantified by cytometry at day 2 (Mean ± SD, n = 3 independent experiments. One-way ANOVA test (two-tailed), at least 10,000 analyzed cells per condition). d % of LOHs (interstitial and terminal) quantified by scSNP-DNAseq (using 5 high-quality SNPs, kilobasic panel in Supplementary Fig. 3f) at day 4 without/with palbociclib exposure. Histogram represents the % of interstitial and telomeric LOH (iLOH in pink and light blue, and tLOH in red and blue). Percentages of LOHs in HSPCs were compared by the Chi-square test (two-tailed). Created in BioRender. Bedel, A. (2025) https://BioRender.com/3fwjwd9.

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