Fig. 8.
AML PDX mice with depleted hCINAP exhibit higher drug sensitivity. a Experimental design for generation of the hCINAP-depleted PDX AML mice models. b, c Representative staining of PB b at day 20 and spleen c at day 26 from AML mice. Scale bars, 50 μm (spleen Ki67, γ-H2AX, and PB TUNEL), 80 μm (splenic white pulp hCINAP). Histograms show the IHC staining quantification of Ki67 + , γ-H2AX+, and hCINAP expression (n = 2 for each group). Expression levels were quantified by the average optical density (AOD) of the positively stained cells with Image-Pro Plus 6.0 in spleens, which were measured by ImageJ. d Representative FACS plots show GFP-positive cells in the bone marrow of mice. n = 2 for each group. e, f Spleens of mice were weighed and photographed at the end of the study. Data were analyzed using Mann–Whitney test. g Kaplan–Meier survival curves of AML mice are shown. hCINAP shRNA, n = 12; Control shRNA, n = 12, ***P < 0.001. h Comparison of the apoptosis rates among white blood cells from AML patients 10 and 11 and control 13 shown in Fig. 1i. Apoptosis analyses were performed and > 105 cells were counted in each group. i Kaplan–Meier analysis of overall survival of patients in the TCGA AML database. The solid and dashed lines represent the low and high expression group, respectively. The difference in the overall survival between these two groups was determined using a log-rank test. j Mice PB cells were collected from the orbit, and the leukocyte were harvested and dissociated. IP assay was done using anti-NPM1 antibody and the NPM1 SUMOylation level was determined by immunoblot using anti-SUMO2/3 antibody. k Quantified NPM1-SUMO PLA score (Left panel) and NPM1-SENP3 PLA score (Right panel) of spleen specimens from the two group of AML mice (n = 2). The representative images are shown in Supplementary Fig. 9a, b. l Scatterplot showing the positive correlation between PLA scores and BRCA1 foci in both AML mice groups. Pearson’s coefficient tests were performed to assess statistical significance. m Working model of hCINAP-mediated chemotherapy and radiotherapy resistance in the DDR. Unprocessed scans of blots are provided in Supplementary Fig. 13