Journal: Nucleus

Article Title: Olivetol induces a non-genotoxic nucleolar DNA damage response via membrane-dependent stress signaling

doi: 10.1080/19491034.2026.2672818

Figure Lengend Snippet: Olivetol induces a nucleolar DNA damage response (n-DDR) without detectable chromosomal DNA breaks. (A) Dose – response curves for Kasumi, MV4;11, KG1, THP1, and HeLa cells treated with increasing concentrations of olivetol for 4 days. Data are presented as mean ± SD ( n = 3 biologically independent experiments). (B) HeLa cells were treated with 10–500 µM olivetol for the indicated time periods or with the DNA topoisomerase II inhibitor etoposide (VP16; 20 µM for 1 h). A neutral comet assay was performed; box plots show the tail moment. Horizontal lines represent the median. **, p < 0.01 by unpaired t-test; n.S., not significant ( n > 500). (C) HeLa cells treated with DMSO (control) or with 50–500 µM olivetol for the indicated time periods were stained for γH2AX (green). DNA was stained with DAPI (blue). (D) HeLa cells were treated with 300 µM olivetol for 30 min and stained for γH2AX (green) and Ki67 (red). (E) HeLa cells were treated with 50–500 µM olivetol for 30 min. ChIP experiments were performed using antibodies against γH2AX. Enriched DNA was analyzed by qPCR with primer pair d1 specific for the rRNA gene promoter or ALU repeats, as indicated in the scheme. Data are presented relative to input. Values represent mean ± SD from at least three independent replicates. **, p < 0.01 by unpaired t-test; n.S., not significant. (F) HeLa cells were treated for 30 min with 300 µM olivetol, 300 µM di-CH3-olivetol (1-butyl-3,5-dimethoxybenzene; BB0282782), 300 µM mono-CH3-olivetol (3-butyl-5-methoxyphenol; BB0282781), 500 µM resveratrol, 500 µM resorcinol, 500 µM quercetin, or 500 µM dihydroquercetin. ChIP experiments were performed using antibodies against γH2AX. Enriched DNA was analyzed by qPCR with primer pair d1 specific for the rRNA gene promoter, as indicated in the scheme. Data are presented relative to input. Values represent mean ± SD from at least three independent replicates. (G) HeLa cells were treated with 300 µM olivetol for 30 min to 3 h and stained for UBF or RPA194 (green) together with NOPP140 or nucleolin (red). DNA was stained with DAPI (blue). (H) HeLa cells were treated with 300 µM olivetol for 30 min and analyzed by transmission electron microscopy (TEM). (I) HeLa cells treated with DMSO (control) or with 300 µM olivetol for 3 h were labeled with 5-ethynyl uridine (EU). EU incorporation was detected by click chemistry. The graph on the right shows quantification of nucleolar EU fluorescence intensity in more than 200 cells. **, p < 0.01 by unpaired t-test.

Article Snippet: Human MV-4–11 (ATCC®CRL-9591TM), THP1 (ATCC®TIB-202TM), KG1 (ATCC®CRL-8031) and Kasumi-1 (ATCC®CRL-2724TM) were cultured in RPMI (PanEco) supplemented with 10% fetal bovine serum (FBS; HyClone/GE Healthcare) and penicillin/streptomycin.

Techniques: Neutral Comet Assay, Control, Staining, Transmission Assay, Electron Microscopy, Labeling, Fluorescence

Journal: Cell Reports Medicine

Article Title: ΔNp73 isoform defines a TP53 -mutant-like poor-risk subgroup of acute myeloid leukemia

doi: 10.1016/j.xcrm.2025.102540

Figure Lengend Snippet: Pharmacological and genetic inhibition of CEBPA synergizes with VEN-induced apoptosis in TP53mut/mut-like AMLs (A) MOLM13 (VEN-sensitive) and KG1 (VEN-resistant) cells were treated for 72 h with increasing concentrations of VEN and GFC. Synergy was determined by Bliss coefficient (ZIP score >10 indicates synergism). (B and C) Drug-induced apoptosis (B) and viable cell counts (C) in MOLM13 shCEBPA/shScr cells treated with VEN alone or in combination with GFC (concentrations indicated in the plots, 72 h) detected by flow cytometry ( n = 4). (D and E) Apoptosis was detected by flow cytometry in gated human CD45 dim CD34 + (or CD117 + cells for CD34 − AMLs) of ex vivo -treated AML samples categorized as TP53 mut-like ( n = 8) (D) and TP53 mut ( n = 9) (E) in a co-culture system using an FITC-annexin V/DAPI staining method. Cells were treated with vehicle, VEN (100 and 500 nM), and VEN+Aza (VEN 100 nM + 5′ Aza 1.5 μM), in the presence or absence of GFC (30 μM) for 72 h. Bar graphs represent the mean ± SEM of all the independent patients screened; each point represents a patient. (F and G) Mitochondrial membrane potential (F) (measured by TMRE staining, n = 18) and total cytoplasmatic ROS levels (G) (measured using the CellROX Red probe, via flow cytometry, n = 6) for the data included in (D) and (E). TP53 mut AMLs are depicted in red, and TP53 mut-like AMLs are depicted in black. APR-246, eprenetapopt. Data are reported as mean ± SEM for (B)–(G). The p values and cell types are indicated in the graphs; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ANOVA and Bonferroni post-test.

Article Snippet: KG1 (male origin) , DSMZ , ACC 14 RRID:CVCL_0374.

Techniques: Inhibition, Flow Cytometry, Ex Vivo, Co-Culture Assay, Staining, Membrane