Fig 2: miR-16, Wip1 and Bcl-2 expression in MCF-7/S and MCF-7/A cells. (A) miR-16 expression was measured by reverse transcription-quantitative PCR in MCF-7/S and MCF-7/A cells. (B) Wip1 and Bcl-2 expression in MCF-7/S and MCF-7/A cells was determined by western blotting. β-actin was the loading control. (C) Co-immunofluorescent staining was applied to measure the expression of Wip1 and Bcl-2 in the indicated cells. Green, red and blue stains represent Wip1, Bcl-2 and the DNA, respectively. Data are presented as the mean ± standard deviation from three independent experiments; **P<0.01. miR, microRNA; Wip1, wild-type p53-induced phosphatase 1.

Fig 3: Blocking WIP1 increases DDR protein phosphorylation and decreases unrepaired DNA damage(A and B) Western blots of (A) hNCC line #1 treated with WIP1 inhibitor (GSK2830371) for 1 h followed by GH (500 ng/mL) for an additional 24 h and (B) organoids treated with WIP1 inhibitor and GH (500 ng/mL) for 48 h. Untreated cells served as control. Representative blots from at least 3 independent experiments are shown. ImageJ quantification of western blots is depicted in Figure S3.(C) Comet assay of hNCC line #1 treated with WIP1 inhibitor for 1 h followed by GH (500 ng/mL) for an additional 24 h. C, control. Single-cell gel electrophoresis was conducted and Olive tail moment assessed on at least 400 cells/per slide for each experiment. Results are shown as mean of 3 experiments ±SEM. Differences were assessed with non-parametric Kruskal-Wallis test followed by Dunn’s multiple comparison test. ∗∗p < 0.01 versus control; ##p < 0.01 GH versus GH + WIP1 inhibitor.

Fig 4: Proposed mechanisms for GH-mediated WIP1 inductionGH activates Src/AMPK phosphorylation. Phospho-AMPK, in turn, phosphorylates HIPK2, which is relocated to the cytoplasm, weakening binding to intranuclear WIP1, decreasing WIP1 ubiquitination, and increasing WIP1 stability. Induced WIP1 dephosphorylates ATM, thus suppressing DDR activity and leading to accumulated DNA damage. Created with BioRender.com.

Fig 5: BST2 promotes IFN–α expression in Wip1−/−MSCs.A, B The mRNA and protein levels of IFN-α in Wip1+/+MSCs and Wip1−/−MSCs were detected by qPCR and ELISA. C Wip1−/− MSCs were transfected with BST2 siRNA, and the mRNA level of BST2 and IFN-α in Wip1−/− MSCs were measured by qPCR 48 h after transfection. Three independent experiments, *P < 0.05, **P < 0.01.